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2024-03-29T15:01:55Z
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http://2012.igem.org/Team:NYMU-Taipei/ymit1
Team:NYMU-Taipei/ymit1
2013-05-08T13:23:13Z
<p>Jayms: Undo revision 300513 by Bert40367 (talk)</p>
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<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymim1" target="_parent">Modeling</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymihpa.html">Human Practice</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
<a href="https://2012.igem.org/Main_Page">iGEM</a><br />
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<div class="title">Team</div><br />
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<p><span class="subtitle">Instructor</span></p><br />
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<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/d/dd/Ymit1.gif" alt="" /> </div><br />
Chuan-Hsiung Chang<br /><br />
Associate Professor<br /><br />
Center for Systems and Synthetic Biology<br /><br />
Institute of Biomedical Informatics<br /><br />
National Yang Ming University<br /><br />
NYMU Synthetic Biology iGEM Team</p><br />
<p><br /><br />
<br /><br />
</p><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/e/ed/Ymit3.gif" alt="" /></div><br />
<p>Chih-Hsien Yang<br /><br />
Ph.D Candidate, Genome Engineering Laboratory, Institute of Biomedical <br /><br />
Informatics, National Yang Ming University, Taiwan</p><br />
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<div id="team_right"><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/b/b2/Ymit2.gif" alt="" /> </div><br />
Pei-Hong Chen<br /><br />
Postdoctoral Fellow<br /><br />
Institute of Bioinformatics &amp; Center for Systems and Synthetic Biology<br /><br />
National Yang-Ming University, Taiwan</p><br />
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<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/a/ab/Ymit4.gif" alt="" /></div><br />
<p>Jesse Wu is a ctrl+s trigger happy person who loves programming.<br /><br />
He likes listening to cantonese pop and Gershwin's Rhapsody in Blue. <br /><br />
In his spare time, he sings. <br /><br />
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Member</span></p><br />
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<div id="team_left"><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/f/f8/T5.gif" alt="" /></div><br />
I am team leader Shi Hao.<br /><br />
My dream is to let the world see Taiwan. <br /><br />
My job in iGEM is to coordinate everything, and make sure that everyone and everything go well.<br />
<p>&nbsp;</p><br />
<p>&nbsp;</p><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/1/17/T7.gif" alt="" width="89" height="125" /></div><br />
<p>I am Chun-Kang.<br /><br />
I am responsible for<br />
contacting entrepreneur iGEM team, implementing Chung Hwa Pulp Corporation project, conducting SRB experiment and propagating iGEM to new NYMU students.</p><br />
<p>&nbsp;</p><br />
<p>&nbsp;</p><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/f/f4/T9.gif" alt="" width="86" height="120" /></div><br />
<p>En-Jie Shih is a second-year student majoring medicine at NYMU. <br /><br />
She is the person in charge of human practice and also helps doing the experiment. </p><br />
<p>&nbsp;</p><br />
<p><br /><br />
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</p><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/2012/4/4b/Ymit11.gif" alt="" width="103" height="132" /></div><br />
<p>Yi Lun is a third-year student specialized in medicine.<br /><br />
He loves movies and soccer.<br />
<br /><br />
He is responsible for Nap gene cloning, modeling and website design.</p><br />
<p>&nbsp;</p><br />
<p>&nbsp;</p><br />
<p>&nbsp;</p><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/1/19/T14.gif" alt="" width="108" height="138" /></div><br />
<p>I&rsquo;m Chris Yen-Chen Lo. My major is Medical technology &amp; Radiology. <br /><br />
I am responsible for sulfur Oxide Terminator Design, gene construction, experiment performance, universal human practice and oral presentation.<br /><br />
</p><br />
</div><br />
<div id="team_right"><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/5/57/T6.gif" alt="" width="97" height="135" /></div><br />
Ting Yi is in the department of Medicine. <br /><br />
He is responsible for most of the cloning experiment and the primary testing design for part BBa-K896000.<br /><br />
As for human practice, he is responsible for the Taiwan iGEMer's fest.<br />
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<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/0/02/T8.gif" alt="" width="91" height="121" /></div><br />
<p>Chiu Hsun-I is a senior in National Yang Ming University majoring in medicine. <br /><br />
Chiu helped with cell culturing, symbiosis experiment and cloning of SQR gene.<br /><br />
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</p><br />
<p>&nbsp;</p><br />
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<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/e/ec/T10.gif" alt="" width="92" height="123" /></div><br />
<p>My name is Chen Shao-Lun. I am in the Department of Medicine. I&rsquo;m responsible for the Cd2+ cleanup subject in our iGEM team. I designed the circuit and did the wet-lab experiment. </p><br />
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<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/b/bb/T12.gif" alt="" width="89" height="120" /></div><br />
<p> Adonis is in medicine department. <br /><br />
Adonis&rsquo; main contribution to NYMU iGEM team includes the design of denitrification, the experiment of Sulfide-quinone Reductase and several human practice. </p><br />
<p>&nbsp;</p><br />
<p><br /><br /><br />
</p><br />
<div class="imgbox" style="margin-left:10px;margin-right:10px;float:left;"><img src="https://static.igem.org/mediawiki/igem.org/5/5b/T15.gif" alt="" width="104" height="134" /></div><br />
<p>Hello, my name is Chung-Yu Huang. I major in Biomedical engineering. <br /><br />
In NYMU-Taipei team, I devoted myself to plan on 2012 Taiwan iGEMer Fest. Also, I participated in many activities which are belong to Human practice part. <br /><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei
Team:NYMU-Taipei
2012-10-27T03:17:37Z
<p>Jayms: </p>
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<a <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymihpa.html">Human Practice</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
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<p><br />
<br />
We planned to cultivate a special strain of genetically engineered cyanobacteria. With reductases metabolizing nitrogen, sulfur and carbon oxides, our organisms reduce three major pollutants in the modern day. Furthermore, we also focus on the removal of cadmium ions from soil.Combining our engineered cyanobacteria and the concept of endosymbiosis, we grant eukaryotes, ultimately human being, the ability to colonize Venus and expand our territory. <br /><br />
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<div class="section_box" align="justify"><br />
<div class="subtitle"><strong>Welcome to our Campus</strong></div><br />
Founded in 1974, National Yang-Ming University, formerly known as National Yang-Ming College of Medicine, has been adhering to the school motto of "benevolent mind and art, putting knowledge into practice" while training excellent humane doctors to provide service by solving medical problems in remote and rural areas.<br /><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei
Team:NYMU-Taipei
2012-10-27T03:15:44Z
<p>Jayms: </p>
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href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
<a href="https://2012.igem.org/Main_Page">iGEM</a><br />
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<br />
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<br />
<br />
<p>&nbsp;</p><br />
<br />
<p><br />
<br />
We planned to cultivate a special strain of genetically engineered cyanobacteria. With reductases metabolizing nitrogen, sulfur and carbon oxides, our organisms reduce three major pollutants in the modern day. Furthermore, we also focus on the removal of cadmium ions from soil.Combining our engineered cyanobacteria and the concept of endosymbiosis, we grant eukaryotes, ultimately human being, the ability to colonize Venus and expand our territory. <br /><br />
<br /><br />
</p><br />
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<li></li><br />
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<li></li><br />
</ul><br />
<div class="section_box" align="justify"><br />
<div class="subtitle"><strong>Welcome to our Campus !</strong></div><br />
Founded in 1974, National Yang-Ming University, formerly known as National Yang-Ming College of Medicine, has been adhering to the school motto of "benevolent mind and art, putting knowledge into practice" while training excellent humane doctors to provide service by solving medical problems in remote and rural areas.<br /><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei
Team:NYMU-Taipei
2012-10-27T03:15:12Z
<p>Jayms: </p>
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href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
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We planned to cultivate a special strain of genetically engineered cyanobacteria. With reductases metabolizing nitrogen, sulfur and carbon oxides, our organisms reduce three major pollutants in the modern day. Furthermore, we also focus on the removal of cadmium ions from soil.Combining our engineered cyanobacteria and the concept of endosymbiosis, we grant eukaryotes, ultimately human being, the ability to colonize Venus and expand our territory. <br /><br />
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<div class="subtitle"><strong>Welcome to our Campus !</strong></div><br />
Founded in 1974, National Yang-Ming University, formerly known as National Yang-Ming College of Medicine, has been adhering to the school motto of "benevolent mind and art, putting knowledge into practice" while training excellent humane doctors to provide service by solving medical problems in remote and rural areas.<br /><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei
Team:NYMU-Taipei
2012-10-27T03:13:32Z
<p>Jayms: </p>
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<p>Founded in 1974, National Yang-Ming University, formerly known as National Yang-Ming College of Medicine, has been adhering to the school motto of &quot;benevolent mind and art, putting knowledge into practice&quot; while training excellent humane doctors to provide service by solving medical problems in remote and rural areas. </p><br />
<p> Our success is no coincidence but an attribute of excellent allocation of limited resources and the devoted efforts of both the faculty and students, all of which were needed to achieve &quot;brilliant results in teaching and research&quot; within such a short period of time. Due to our research atmosphere, academic accomplishments, innovative knowledge and cultivation of talent, we have well demonstrated our standing as a top university. <br /><br />
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<div class="subtitle"><strong>Welcome to our Campus !</strong></div><br />
Founded in 1974, National Yang-Ming University, formerly known as National Yang-Ming College of Medicine, has been adhering to the school motto of "benevolent mind and art, putting knowledge into practice" while training excellent humane doctors to provide service by solving medical problems in remote and rural areas.<br />
Our success is no coincidence but an attribute of excellent allocation of limited resources and the devoted efforts of both the faculty and students, all of which were needed to achieve "brilliant results in teaching and research" within such a short period of time. Due to our research atmosphere, academic accomplishments, innovative knowledge and cultivation of talent, we have well demonstrated our standing as a top university. <br />
<br />
<br />
<br />
<br />
We planned to cultivate a special strain of genetically engineered cyanobacteria. With reductases metabolizing nitrogen, sulfur and carbon oxides, our organisms reduce three major pollutants in the modern day. Furthermore, we also focus on the removal of cadmium ions from soil.Combining our engineered cyanobacteria and the concept of endosymbiosis, we grant eukaryotes, ultimately human being, the ability to colonize Venus and expand our territory. <br /><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei
Team:NYMU-Taipei
2012-10-27T03:13:11Z
<p>Jayms: </p>
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<br />
<br />
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<p>Founded in 1974, National Yang-Ming University, formerly known as National Yang-Ming College of Medicine, has been adhering to the school motto of &quot;benevolent mind and art, putting knowledge into practice&quot; while training excellent humane doctors to provide service by solving medical problems in remote and rural areas. </p><br />
<p> Our success is no coincidence but an attribute of excellent allocation of limited resources and the devoted efforts of both the faculty and students, all of which were needed to achieve &quot;brilliant results in teaching and research&quot; within such a short period of time. Due to our research atmosphere, academic accomplishments, innovative knowledge and cultivation of talent, we have well demonstrated our standing as a top university. <br /><br />
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We planned to cultivate a special strain of genetically engineered cyanobacteria. With reductases metabolizing nitrogen, sulfur and carbon oxides, our organisms reduce three major pollutants in the modern day. Furthermore, we also focus on the removal of cadmium ions from soil.Combining our engineered cyanobacteria and the concept of endosymbiosis, we grant eukaryotes, ultimately human being, the ability to colonize Venus and expand our territory. <br /><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymiq5.html
Team:NYMU-Taipei/ymiq5.html
2012-10-27T03:12:32Z
<p>Jayms: </p>
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<div class="title">Further Experiments after Asia Jamboree</div><br />
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<br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/0/0a/Ymiqw1.png" alt="" width="502" height="333" /><br /><br />
<p align="center">Sulfite quinine reductase, SQR use hydrogen sulfide, H2S as electron resource to perform photosynthesis<br /><br />
</p></div><br />
In previous experiment, we’ve demonstrated that Sulfide-Quinone Reductase (sqr) expressed in Synechococcus sp. PCC7942 enhanced the growth of the unicellular cyanobacteria. Nevertheless, most of our observations were based on the difference of optical density between wild type and sqr expressing strain. Though it is quite common to estimate the growth of cyanobacteria by measuring the optical density under certain wavelength, such measurement only provides an approximate data for comparison, and various factors can influence the observation accuracy, With regard to that, we conduct another series of experiments via flow cytometer and plate count method to examine the exact cell concentration. <br />
<br /><br />
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<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/f/f2/Ymiqw2.png" alt="" width="398" height="314" /><br /><br />
Cyanobacteria contain different of cytochrome. We can detect them by different wavelength fluorescent light.<br /><br />
( http://www.masa.asn.au/phpBB3/viewtopic.php?t=233828&amp;p=960435 )<br /><br />
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<br /><div align="left"><br />
<p><span class="subtitle">Cyanobacteria Culture </span></p><br />
</div><br />
<p>Both wild type and sqr stain of <em>Synechococcus sp. PCC7942</em> were cultivated in BG-11 medium (Allen M.M. 1968) [1]. The initial cell concentration was adjusted to an OD730 of 0.2. The experiment was initialized after the addition of unicellular blue-green algae, and the six-well plates were placed on a 100 r.p.m. shaker.<br />
<br /><br />
</p><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/c/ce/Ymiqw3.png" alt="" width="434" height="334" /><br /><br />
We culture Synechococcus sp. PCC7942 wildtype and sqr transformed one in BG-11 medium with suitable shaker. <br /><br />
<br /><br />
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<br /><div align="left"><br />
<p><span class="subtitle">DCMU blocking assay</span></p><br />
</div><br />
<br />
<br />
<p>Freshly made sodium sulfide containing medium was added into the experiment group. The concentration of sodium sulfide was 2.5 mM, and additional 2.5mM of sodium sulfide was added every 24 hours. 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) [2], a chemical which can block photosystem II was also added into both experiment and control group. A positive control group was established by simply cultivated cyanobacteria in fresh medium. As followed, we analyze by flow cytometry analysis, 730 nm absorption assay and plating of bacteria.<br />
<br /><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/2/23/Ymiqw4.png" alt="" width="558" height="262" /><br /><br />
DCMU specifically block photosynthesis system II<br /><br />
( http://5e.plantphys.net/article.php?ch=7&id=75 )<br /><br />
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<br /><br />
</div><br />
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<p><span class="subtitle">Flow cytometry analysis </span></p><br />
</div><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/2/28/Ymiqw5.png" alt="" width="475" height="361" /><br /><br />
We use flow cytometry with multiple wavelength fluorescent light to find out the differences between two groups. <br /><br />
<br /><br />
</div><br />
<p>The automation of viability analysis method via flow cytometer was reported in previous research [3]. In our experiment, cells were analyzed using a flow cytometer FACSCanto. The chlorophyll fluorescence filter set (excitation: 435/40 nm; beam splitter: 510 nm; emission: 515 nm long-pass) were used [3]. By using the automated counting function of FACSCanto, we examined the accurate number of cell. The green fluorescence of the cell area was constrained at an intensity value of 50 to differentiate between red (viable) and green (non-viable) cells [3]. The percentage of red and green cells and the exact cell concentration was calculated.<br />
</p><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/3/3e/Ymiqw6.png" alt="" width="558" height="385" /><br /><br />
Different fluorescent dyes and its wavelength<br />
(http://www.epitomics.com/images/DyLight%20Spectra%20Chart.jpg )<br />
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<div class="title">Results &amp; Discussion</div><br />
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<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/f/f9/Ymiqw7.png" width="600" height="300" /><br /><br />
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<br /><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/b/ba/Ymiqw8.png" width="600" height="283" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/f/fc/Ymiqw9.png" width="600" height="262" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/d/d5/Ymiqw10.png" width="600" height="266" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/b/b1/Ymiqw11.png" width="600" height="291" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/7/72/Ymiqw12.png" width="600" height="249" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/b/bc/Ymiqw13.png" width="600" height="244" /><br /><br />
</div><br />
<br /><br />
<ol><br />
<li>We kill wildtype cyanobacteria with 4oC environment for 2 weeks</li><br />
<li>We find out that when adding Na2S to our DCMU blocked SQR bacteria, they show more fluorescent light(higher photosynthesis ability, live better)</li><br />
<li>We find out that when bacteria are challenged with DCMU, SQR group can live better than Wldtype group, which means SQR gene help cyanobacteria survive in the adversity(DCMU, Na2S)</li><br />
<li>It&rsquo;s very interesting when we compare data:</li><br />
</ol><br />
<br /><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/3/3a/Ymiqw14.png" alt="" width="422" height="302" /><br /><br />
<p align="left">When DCMU challenge bacteria, SQR transformed croup can produce more fluorescent light(higher photosynthesis ability, live better)</div><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/6/6d/Ymiqw15.png" alt="" width="405" height="299" /><br />
<p align="left">In SQR group, we use DCMU to challenge bacteria. Adding Na2S bacteria produce more fluorescent light. </div><div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/b/b9/Ymiqw16.png" alt="" width="382" height="280" /><br />
<p align="left">We survey several papers and hypothesize that the fluorescent comes from cytochrome b6f, which is an important role in the downstream of photosynthesis system II and the electron transport chain. (Erwin J. G. Peterman,Stephan-Olav Wenk et al, Fluorescence and Absorption Spectroscopy of the Weakly Fluorescent Chlorophyll a in Cytochrome b6f of Synechocystis PCC6803, Biophysical Journal Volume 75 July 1998 389–398)<br /><br />
<br /><br />
<br /><br />
</div><br />
<div align="left"><br />
<p><span class="subtitle">730 nm absorption</span></p><br />
</div><br />
<p>Based on the flow cytometry result, we double check our bacteria amount by 730nm absorption:</p><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/9/9f/Ymiqw17.png" alt="" width="559" height="394" /><br /><br />
</div><br />
<ol><br />
<li>When challenge bacteria with DCMU, SQR transformed cyanobacteria live better than wildtype(higher photosynthesis, better adversity resistance)</li><br />
<li>It&rsquo;s very interesting that when add Na2S into two kinds of bacteria, it might produce more difficulty for living(maybe the Na2S concentration is wrong? HS- binds to Fe and generate worse environment for bacteria)<br /><br />
<br /><br />
</li><br />
</ol><br />
<div align="left"><br />
<p><span class="subtitle">Plating of bacteria</span></p><br />
</div><br />
<p>Viable cell concentrations were also determined by spreading 0.1 mL of culturing cyanobacteria on BG-11 agar plates. Colony counts were conducted after these plates were incubated for 7 days at room temperature. The plating was performed in duplicates for every sample.</p><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/6/6c/Ymiqw18.png" width="509" height="557" /><br /><br />
<p align="left" >We culture bacteria into solid plate. After incubation for about 10 days, we will count the colonies and compare each group. <br /><br />
<br /><br />
<br /><br />
</div><br />
<div align="left"><br />
<p><span class="subtitle">CO2 consumption analysis</span><br /><br />
</p><br />
</div><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/2/25/Ymiqw19.png" alt="" width="407" height="482" /><br /><br />
<p align="left" >We culture two group bacteria with stable lamina CO2 input and compare the CO2 for each group.<br /><br />
</div><br />
<p>It&rsquo;s very interesting that when adding Na2S to both two bacteria group, we get the similar result. We hypothesize that Na2S might generate adversity to bacteria. Thus, It&rsquo;s very important to analyze CO2 consumption. CO2 concentration and analysis were examined by the infrared CO2 analyzer and FL22 Algal CO2 Package respectively [4]. Cyanobacteria samples were cultured for 24 hours following CO2 analysis. CO2 fixation rates were calculated in terms of CO2 exchange rate as the following equation: </p><br />
<div class=out style='text-align:center'><br />
<p align="center" ><img src="https://static.igem.org/mediawiki/2012/d/dd/Ymiqw20.png" alt="" width="433" height="60" /> </div><br />
<p>The HCO3 concentration in the culture medium was also measured with Carbon Dioxide Enzymatic Assay of BQ Kits (Bio-Quant, USA).</p><br />
<div class=out style='text-align:center'><br />
<p align="center" ><img src="https://static.igem.org/mediawiki/2012/b/bc/Ymiqw21.png" alt="" width="558" height="320" /> <br /><br />
<p align="left"> We will analyze CO2 consumption for SQR grop and wildtype cyanobacteria to find out the activity of them. Finally, We will use DCMU to challenge two groups of bacteria and perform this assay again ( http://www.quora.com/Carbon-Dioxide ) <br /><br />
<br /><br />
<br /><br />
</div><div align="left"><br />
<p><span class="subtitle">References</span><br /><br />
</p><br />
</div><br />
<ol><br />
<li><em>Allen MM (1968) Simple conditions for growth of unicellular blue-green algae. J Phycol 4: 1-3</em></li><br />
<li><em>Adaptation to Hydrogen Sulfide of Oxygenic and Anoxygenic Photosynthesis among Cyanobacteria YEHUDA COHEN,'* BO BARKER J0RGENSEN,2 NIELS PETER REVSBECH,2 AND RICARDO POPLAWSKIl H. Steinitz Marine Biology Laboratory, The Hebrew University of Jerusalem, Eilat 88103, Israel,1 and Institute of Ecology and Genetics, University of Aarhus, Ny Munkegade, DK 8000, Aarhus C, Denmark2 Received 1 July 1985/Accepted 22 October 1985</em></li><br />
<li><em>A simple viability analysis for unicellular cyanobacteria using a new autofluorescence assay, automated microscopy, and ImageJ Katja Schulze1,2*, Diana A López1, Ulrich M Tillich1 and Marcus Frohme1 BMC Biotechnology 2011, 11:118 doi:10.1186/1472-6750-11-118</em></li><br />
<li><em>Enhancing CO2 bio-mitigation by genetic engineering of cyanobacteria† Pei-Hong Chen,a Hsien-Lin Liu,a Yin-Ju Chen,a Yi-Hsiang Cheng,a Wei-Ling Lin,a Chien-Hung Yeha and Chuan-Hsiung Chang*ab Received 16th January 2012, Accepted 7th June 2012 DOI: 10.1039/c2ee21124f<br /><br />
</em> <br /><br />
</li><br />
</ol><br />
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<h2 class="drawer-handle open">Sulfide as Energy Generator</h2><br />
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</ul><br />
</li><br />
<br />
<br />
<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
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<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
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References</a></li><br />
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Jayms
http://2012.igem.org/File:Ymiqw21.png
File:Ymiqw21.png
2012-10-27T03:12:20Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw20.png
File:Ymiqw20.png
2012-10-27T03:11:59Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw19.png
File:Ymiqw19.png
2012-10-27T03:11:41Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw18.png
File:Ymiqw18.png
2012-10-27T03:11:19Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw17.png
File:Ymiqw17.png
2012-10-27T03:11:00Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw16.png
File:Ymiqw16.png
2012-10-27T03:10:43Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw15.png
File:Ymiqw15.png
2012-10-27T03:10:24Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw14.png
File:Ymiqw14.png
2012-10-27T03:09:56Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw13.png
File:Ymiqw13.png
2012-10-27T03:09:37Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw12.png
File:Ymiqw12.png
2012-10-27T03:09:17Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw11.png
File:Ymiqw11.png
2012-10-27T03:08:48Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw10.png
File:Ymiqw10.png
2012-10-27T03:08:17Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw9.png
File:Ymiqw9.png
2012-10-27T03:07:40Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw8.png
File:Ymiqw8.png
2012-10-27T03:07:18Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw7.png
File:Ymiqw7.png
2012-10-27T03:06:56Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw6.png
File:Ymiqw6.png
2012-10-27T03:06:35Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw5.png
File:Ymiqw5.png
2012-10-27T03:06:11Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw4.png
File:Ymiqw4.png
2012-10-27T03:05:44Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw3.png
File:Ymiqw3.png
2012-10-27T03:05:20Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw2.png
File:Ymiqw2.png
2012-10-27T03:04:55Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/File:Ymiqw1.png
File:Ymiqw1.png
2012-10-27T03:04:18Z
<p>Jayms: </p>
<hr />
<div></div>
Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymiq5.html
Team:NYMU-Taipei/ymiq5.html
2012-10-27T03:01:20Z
<p>Jayms: </p>
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<div class="title">Further Experiments after Asia Jamboree</div><br />
<br /><br />
<div align="left"><br />
<br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw1.png" alt="" width="502" height="333" /><br /><br />
<p align="center">Sulfite quinine reductase, SQR use hydrogen sulfide, H2S as electron resource to perform photosynthesis<br /><br />
</p></div><br />
In previous experiment, we’ve demonstrated that Sulfide-Quinone Reductase (sqr) expressed in Synechococcus sp. PCC7942 enhanced the growth of the unicellular cyanobacteria. Nevertheless, most of our observations were based on the difference of optical density between wild type and sqr expressing strain. Though it is quite common to estimate the growth of cyanobacteria by measuring the optical density under certain wavelength, such measurement only provides an approximate data for comparison, and various factors can influence the observation accuracy, With regard to that, we conduct another series of experiments via flow cytometer and plate count method to examine the exact cell concentration. <br />
<br /><br />
<br /><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw2.png" alt="" width="398" height="314" /><br /><br />
Cyanobacteria contain different of cytochrome. We can detect them by different wavelength fluorescent light.<br /><br />
( http://www.masa.asn.au/phpBB3/viewtopic.php?t=233828&amp;p=960435 )<br /><br />
<br /><br />
<br /><br />
</div><br />
<br /><div align="left"><br />
<p><span class="subtitle">Cyanobacteria Culture </span></p><br />
</div><br />
<p>Both wild type and sqr stain of <em>Synechococcus sp. PCC7942</em> were cultivated in BG-11 medium (Allen M.M. 1968) [1]. The initial cell concentration was adjusted to an OD730 of 0.2. The experiment was initialized after the addition of unicellular blue-green algae, and the six-well plates were placed on a 100 r.p.m. shaker.<br />
<br /><br />
</p><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw3.png" alt="" width="434" height="334" /><br /><br />
We culture Synechococcus sp. PCC7942 wildtype and sqr transformed one in BG-11 medium with suitable shaker. <br /><br />
<br /><br />
</div><br />
<br /><div align="left"><br />
<p><span class="subtitle">DCMU blocking assay</span></p><br />
</div><br />
<br />
<br />
<p>Freshly made sodium sulfide containing medium was added into the experiment group. The concentration of sodium sulfide was 2.5 mM, and additional 2.5mM of sodium sulfide was added every 24 hours. 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) [2], a chemical which can block photosystem II was also added into both experiment and control group. A positive control group was established by simply cultivated cyanobacteria in fresh medium. As followed, we analyze by flow cytometry analysis, 730 nm absorption assay and plating of bacteria.<br />
<br /><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw4.png" alt="" width="558" height="262" /><br /><br />
DCMU specifically block photosynthesis system II<br /><br />
( http://5e.plantphys.net/article.php?ch=7&id=75 )<br /><br />
<br /><br />
<br /><br />
</div><br />
<br /><br />
<div align="left"><br />
<p><span class="subtitle">Flow cytometry analysis </span></p><br />
</div><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw5.png" alt="" width="475" height="361" /><br /><br />
We use flow cytometry with multiple wavelength fluorescent light to find out the differences between two groups. <br /><br />
<br /><br />
</div><br />
<p>The automation of viability analysis method via flow cytometer was reported in previous research [3]. In our experiment, cells were analyzed using a flow cytometer FACSCanto. The chlorophyll fluorescence filter set (excitation: 435/40 nm; beam splitter: 510 nm; emission: 515 nm long-pass) were used [3]. By using the automated counting function of FACSCanto, we examined the accurate number of cell. The green fluorescence of the cell area was constrained at an intensity value of 50 to differentiate between red (viable) and green (non-viable) cells [3]. The percentage of red and green cells and the exact cell concentration was calculated.<br />
</p><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw6.png" alt="" width="558" height="385" /><br /><br />
Different fluorescent dyes and its wavelength<br />
(http://www.epitomics.com/images/DyLight%20Spectra%20Chart.jpg )<br />
</div><br />
<br /><br />
<br /><br />
<br /><br />
<div class="title">Results &amp; Discussion</div><br />
<br /><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw7.png" width="600" height="300" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw8.png" width="600" height="283" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw9.png" width="600" height="262" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw10.png" width="600" height="266" /><br /><br />
</div><br />
<br /><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw11.png" width="600" height="291" /><br /><br />
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<br /><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw12.png" width="600" height="249" /><br /><br />
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<br /><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw13.png" width="600" height="244" /><br /><br />
</div><br />
<br /><br />
<ol><br />
<li>We kill wildtype cyanobacteria with 4oC environment for 2 weeks</li><br />
<li>We find out that when adding Na2S to our DCMU blocked SQR bacteria, they show more fluorescent light(higher photosynthesis ability, live better)</li><br />
<li>We find out that when bacteria are challenged with DCMU, SQR group can live better than Wldtype group, which means SQR gene help cyanobacteria survive in the adversity(DCMU, Na2S)</li><br />
<li>It&rsquo;s very interesting when we compare data:</li><br />
</ol><br />
<br /><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw14.png" alt="" width="422" height="302" /><br /><br />
<p align="left">When DCMU challenge bacteria, SQR transformed croup can produce more fluorescent light(higher photosynthesis ability, live better)</div><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw15.png" alt="" width="405" height="299" /><br />
<p align="left">In SQR group, we use DCMU to challenge bacteria. Adding Na2S bacteria produce more fluorescent light. </div><div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw16.png" alt="" width="382" height="280" /><br />
<p align="left">We survey several papers and hypothesize that the fluorescent comes from cytochrome b6f, which is an important role in the downstream of photosynthesis system II and the electron transport chain. (Erwin J. G. Peterman,Stephan-Olav Wenk et al, Fluorescence and Absorption Spectroscopy of the Weakly Fluorescent Chlorophyll a in Cytochrome b6f of Synechocystis PCC6803, Biophysical Journal Volume 75 July 1998 389–398)<br /><br />
<br /><br />
<br /><br />
</div><br />
<div align="left"><br />
<p><span class="subtitle">730 nm absorption</span></p><br />
</div><br />
<p>Based on the flow cytometry result, we double check our bacteria amount by 730nm absorption:</p><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw17.png" alt="" width="559" height="394" /><br /><br />
</div><br />
<ol><br />
<li>When challenge bacteria with DCMU, SQR transformed cyanobacteria live better than wildtype(higher photosynthesis, better adversity resistance)</li><br />
<li>It&rsquo;s very interesting that when add Na2S into two kinds of bacteria, it might produce more difficulty for living(maybe the Na2S concentration is wrong? HS- binds to Fe and generate worse environment for bacteria)<br /><br />
<br /><br />
</li><br />
</ol><br />
<div align="left"><br />
<p><span class="subtitle">Plating of bacteria</span></p><br />
</div><br />
<p>Viable cell concentrations were also determined by spreading 0.1 mL of culturing cyanobacteria on BG-11 agar plates. Colony counts were conducted after these plates were incubated for 7 days at room temperature. The plating was performed in duplicates for every sample.</p><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw18.png" width="509" height="557" /><br /><br />
<p align="left" >We culture bacteria into solid plate. After incubation for about 10 days, we will count the colonies and compare each group. <br /><br />
<br /><br />
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</div><br />
<div align="left"><br />
<p><span class="subtitle">CO2 consumption analysis</span><br /><br />
</p><br />
</div><br />
<div class=out style='text-align:center'><img src="../../../Users/jaymes/Desktop/ymiqw19.png" alt="" width="407" height="482" /><br /><br />
<p align="left" >We culture two group bacteria with stable lamina CO2 input and compare the CO2 for each group.<br /><br />
</div><br />
<p>It&rsquo;s very interesting that when adding Na2S to both two bacteria group, we get the similar result. We hypothesize that Na2S might generate adversity to bacteria. Thus, It&rsquo;s very important to analyze CO2 consumption. CO2 concentration and analysis were examined by the infrared CO2 analyzer and FL22 Algal CO2 Package respectively [4]. Cyanobacteria samples were cultured for 24 hours following CO2 analysis. CO2 fixation rates were calculated in terms of CO2 exchange rate as the following equation: </p><br />
<div class=out style='text-align:center'><br />
<p align="center" ><img src="../../../Users/jaymes/Desktop/ymiqw20.png" alt="" width="433" height="60" /> </div><br />
<p>The HCO3 concentration in the culture medium was also measured with Carbon Dioxide Enzymatic Assay of BQ Kits (Bio-Quant, USA).</p><br />
<div class=out style='text-align:center'><br />
<p align="center" ><img src="../../../Users/jaymes/Desktop/ymiqw21.png" alt="" width="558" height="320" /> <br /><br />
<p align="left"> We will analyze CO2 consumption for SQR grop and wildtype cyanobacteria to find out the activity of them. Finally, We will use DCMU to challenge two groups of bacteria and perform this assay again ( http://www.quora.com/Carbon-Dioxide ) <br /><br />
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<br /><br />
</div><div align="left"><br />
<p><span class="subtitle">References</span><br /><br />
</p><br />
</div><br />
<ol><br />
<li><em>Allen MM (1968) Simple conditions for growth of unicellular blue-green algae. J Phycol 4: 1-3</em></li><br />
<li><em>Adaptation to Hydrogen Sulfide of Oxygenic and Anoxygenic Photosynthesis among Cyanobacteria YEHUDA COHEN,'* BO BARKER J0RGENSEN,2 NIELS PETER REVSBECH,2 AND RICARDO POPLAWSKIl H. Steinitz Marine Biology Laboratory, The Hebrew University of Jerusalem, Eilat 88103, Israel,1 and Institute of Ecology and Genetics, University of Aarhus, Ny Munkegade, DK 8000, Aarhus C, Denmark2 Received 1 July 1985/Accepted 22 October 1985</em></li><br />
<li><em>A simple viability analysis for unicellular cyanobacteria using a new autofluorescence assay, automated microscopy, and ImageJ Katja Schulze1,2*, Diana A López1, Ulrich M Tillich1 and Marcus Frohme1 BMC Biotechnology 2011, 11:118 doi:10.1186/1472-6750-11-118</em></li><br />
<li><em>Enhancing CO2 bio-mitigation by genetic engineering of cyanobacteria† Pei-Hong Chen,a Hsien-Lin Liu,a Yin-Ju Chen,a Yi-Hsiang Cheng,a Wei-Ling Lin,a Chien-Hung Yeha and Chuan-Hsiung Chang*ab Received 16th January 2012, Accepted 7th June 2012 DOI: 10.1039/c2ee21124f<br /><br />
</em> <br /><br />
</li><br />
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<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<li class="drawer"><br />
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<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymis8.html">Further Experiments after Asia Jamboree</a></li><br />
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<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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</li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymiq5.html
Team:NYMU-Taipei/ymiq5.html
2012-10-27T01:52:17Z
<p>Jayms: Created page with "<html xmlns="http://www.w3.org/1999/xhtml"> <head> <meta http-equiv="Content-Type" content="text/html; charset=utf-8" /> <title>NYMU iGEM</title> <link href="http://www.royals.co..."</p>
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<div class="title">Methods</div><br />
<div align="left"><br />
<p><span class="subtitle">Resistance of Cyanobacteria (Synechococcus SP. PCC 7002) to Sulfide compound</span></p><br />
</div><br />
<div align="left"><br />
<p>Several Cyanobacteria have Sulfide-Quinone Reductase (sqr) and thus the ability to deprive electron from sulfide compound. According to both databases of NCBI and KEGG, the sqr in Synechococcus SP. PCC 7002 shared great similarity with that of Oscillatoria limnetica, which is reported to exhibit anoxygenic photosynthesis by consumed sulfide anion. Since we planned to express sqr from Synechococcus SP. PCC 7002 in Synechococcus SP. PCC 7942 and Escherichia coli, the experiment was designed to testify the property of the sqr. DCMU was added in the medium to inhibit photosystem II, and therefore only sodium sulfide in the medium can provide electron for carbon photoassimilation. By creating different dilution of sodium sulfide, we expected that the more sodium sulfide was present, the better the cell grew. <br /><br />
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<br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/5/5e/Ymiq3.png" width="469" height="276" border="0" align="center" /><br /><br />
DCMU structure and its mechanism on photosynthesis<br /><br />
http://en.wikipedia.org/wiki/File:Diuron.png <br /><br />
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<div align="left"><br />
<p><span class="subtitle">Sodium sulfide concentration and cell growth</span></p><br />
</div><br />
<p>From the previous studies, it is suggested that <em>Synechococcus SP. PCC 7002 </em>is able to metabolize sulfide compounds. We took advantage of the results in our last experiment and adjusted the concentration of DCMU to an appropriate degree. Since sulfide would become the main reducing energy for photoassimilation under the effect of DCMU, we believe the more sulfide concentration in the wells, the better cell growth would be observed.</p><br />
<div class=out style='text-align:center'><span class="out" style="text-align:center"><img src="https://static.igem.org/mediawiki/2012/2/23/Ymiq4-1.png" width="557" height="415" border="0" align="center" /></span><br /><br />
<br /><br />
<br />
<br />
<p><span class="subtitle">The effect of sodium sulfide on Synechococcus SP. PCC 7942 growth rate</span></p><br />
</div><br />
<br />
<p>After thoroughly examined the ability of sqr in Synechococcus SP. PCC 7002, we planned to conduct a series of similar experiments on Synechococcus SP. PCC 7942. Except for the cultivation medium, other growing conditions remained the same. Instinctively, the strain expressing sqr should grow better than the wile type strain. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper.</p><br />
<div class=out style='text-align:center'><span class="out" style="text-align:center"><img src="https://static.igem.org/mediawiki/2012/5/57/Ymiq4-2.png" width="573" height="299" border="0" align="center" /></span><br /><br />
<div align="left"><br />
<p><span class="subtitle">Sulfide concentration and the growth of sqr expressing strain Synechococcus SP. PCC 7942</span></p><br />
</div><br />
<div align="left"><br />
<p>It was expected that SQR expressing strain and wild type counterpart would have different growth rate under the presence of sulfide compounds. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper. As the result, we analyze H2S amount to detect whether sqr gene work or not. Therefore, we perform Chemical microvolume turbidimetry method to detect H2S concentration (see Sulfur Oxide Terminator part)</div><br /><br />
<br />
<div align="left"><br />
<p><span class="subtitle">Sulfide oxidation in Escherichia coli expressing sulfide-quinone reductase gene</span></p><br />
</div><br />
<div align="left"><br />
<p>Repots have it that Escherichia coli can express functional sulfide-quinone reductase (SQR). Therefore, we slightly adjusted the previous experiment and applied to the SQR gene from Synechococcus SP. PCC 7002. With methylene blue method, we would test the efficiency of SQR sulfide oxidation. Since such method involved in measurement of optical density, it is more appropriate to perform such experiment on colorless bacteria instead of engineered cyanobacteria strain.</div><br /><br />
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<h2 class="drawer-handle open">Sulfide as Energy Generator</h2><br />
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<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymis8.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html
Team:NYMU-Taipei/ymivenusiancr.html
2012-10-27T01:51:48Z
<p>Jayms: </p>
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<div class="title">Conclusion &amp; References</div><br />
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<p>Inv-llo cyanobacteria coculturing with induced pluripotent stem cells from mice, and J774 mouse macrophage cell line separately suggest the symbiosis possibility by the recruitment of cyanobacteria and the faster entry and escape rate.</p><br />
<p><br /><br />
References<br /><br />
</p><br />
<p>1. The Pore-Forming Toxin Listeriolysin O Mediates a Novel Entry Pathway of L. monocytogenes into Human Hepatocytes, Stephen Vadia1, Eusondia Arnett1, Anne-Cécile Haghighat1, Elisabeth M. Wilson-Kubalek2, Rodney K. Tweten3, Stephanie Seveau1*, PLoS One. 2011; 7(11): e1002356</p><br />
<p>2. Towards a Synthetic Chloroplast, Christina M. Agapakis,1 Henrike Niederholtmeyer,1,¤ Ramil R. Noche,1 Tami D. Lieberman,1 Sean G. Megason,1 Jeffrey C. Way,2 and Pamela A. Silver1,2,PLoS One. 2011; 6(4): e18877.<br /><br />
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<h2 class="drawer-handle open">Becoming Venusian</h2><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
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<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
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<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
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<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
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<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
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<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Result</a>s &amp; References</li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
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<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
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<h2 class="drawer-handle">Cd+2 Collector</h2><br />
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<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
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<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html
Team:NYMU-Taipei/ymivenusianrd.html
2012-10-27T01:51:39Z
<p>Jayms: </p>
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<div class="title">Result &amp; Discussion</div><br />
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<p>There were three different types of cells we chose to do the coculture experiment. Part 1 revealed that J774 macrophage cell line enabled the entry of cyanobacteria by phagocytosis. Part2 and part3 were both mouse induced pluripotent stem cells but in different stage of stem cells. Secondly was non-EB iPS cells, separated from mouse embryonic fibroblasts (MEFs) and subcultured on culture dish. Thirdly was iPS cells in the Embryoid body (EB) stage. Later experiments showed three parts of results, J774 macrophage cell line(figure 2), non-EB iPS cells(figure 3), and iPS cells at the EB stage(figure 4). Red pots were S. elongates PCC7942 under fluorescence microscope 530-550 nm excitation filter. Under the transmitted light, transparent one was cells. </p><br />
<p>Part 1 <br /><br />
<br /><br />
Figure 2 revealed coculturing results of J774 macrophage cell lines and S. elongatus PCC 7942(wildtype/inv-llo). After 24-hour coculturing and 24-hour cell culture under gentamycin 100 ug/ml, same concentration of wildtype and inv-llo transformed type(both were OD 0.05) both entered into J774 macrophage cell line by phagocytosis and the help of inv-llo.</p><br />
<table border="1" cellspacing="0" cellpadding="0" width="634"><br />
<tr><br />
<td width="313" valign="top"><p><img width="298" height="224" src="https://static.igem.org/mediawiki/igem.org/7/74/Ymivenusianrd_clip_image002.jpg" /> <br /><br />
(A)inv-llo strain, merged image of fluorescence and bright field</p></td><br />
<td width="321" valign="top"><p><img width="306" height="230" src="https://static.igem.org/mediawiki/igem.org/7/71/Ymivenusianrd_clip_image004.jpg" /> <br /><br />
(B) inv-llo strain, image of fluorescence </p></td><br />
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<td width="313" valign="top"><p><img width="298" height="224" src="https://static.igem.org/mediawiki/igem.org/8/82/Ymivenusianrd_clip_image006.jpg" /> <br /><br />
(C) wildtype, merged image of fluorescence and bright field</p></td><br />
<td width="321" valign="top"><p><img width="296" height="222" src="https://static.igem.org/mediawiki/igem.org/2/2c/Ymivenusianrd_clip_image008.jpg" /> <br /><br />
(D) wildtype, image of fluorescence</p></td><br />
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<tr><br />
<td width="634" colspan="2" valign="top"><p>Figure 2. Red pots were S. elongates PCC7942. Round and transparent one was J774 macrophage cell line. By phagocytosis, wildtype and inv-llo type entered into cells. </p></td><br />
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</table><br />
<p>In figure 2,(B)(D) were the images under the fluorescence microscope 530-550 nm excitation filter. Red pots were S. elongates PCC7942. Under the transmitted light, round and transparent one was J774 macrophage cell line. (A)(C) were the merged images of bright fields and fluorescence. Compared inv-llo transformed type to wildtype, red pots of inv-llo transformed type were less bright than those of wildtype. Additionally, inv-llo transformed type numbers in cells were less than wildtype numbers. It might suggest that inv-llo gene could raise the entry speed and the escape rate. </p><br />
<p>Part 2<br /><br />
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Figure 3 showed the coculture results of non-EB iPS cells. Cells and S.elongatus PCC7942(wildtype/inv-llo). Images(A)(B)(E)(F)(G)(H) were inv-llo strain and non-EB iPS cells coculturing results (A,B: under fluorescence microscope E~H: under confocal laser microscope.) Cells and cyanobacteria at the separate focus revealed the negative symbiosis. However, inv-llo plasmid showed that inv-llo would recruit cyanobacteria to cells. 3D image under the confocal laser microscope(image H) revealed the recruitment of cyanobacteria.</p><br />
<table border="1" cellspacing="0" cellpadding="0" width="614"><br />
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<td width="318" valign="top"><p><img width="273" height="204" src="https://static.igem.org/mediawiki/igem.org/7/75/Ymivenusianrd_clip_image002_0000.jpg" /> <br /><br />
(A) inv-llo strain, merged image of fluorescence and bright field</p></td><br />
<td width="296" valign="top"><p><img width="279" height="209" src="https://static.igem.org/mediawiki/igem.org/b/bb/Ymivenusianrd_clip_image004_0000.jpg"/> <br /><br />
(B) inv-llo strain, image of fluorescence</p></td><br />
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<td width="318" valign="top"><p><img width="273" height="204" src="https://static.igem.org/mediawiki/igem.org/1/1e/Ymivenusianrd_clip_image006_0000.jpg" /> <br /><br />
(C) wildtype, merged image of fluorescence and bright field</p></td><br />
<td width="296" valign="top"><p><img width="267" height="200" src="https://static.igem.org/mediawiki/igem.org/b/bb/Ymivenusianrd_clip_image008_0000.jpg<br />
"/> <br /><br />
(D) wildtype, image of fluorescence</p></td><br />
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<tr><br />
<td width="318" valign="top"><p><img src="https://static.igem.org/mediawiki/igem.org/b/bb/Ymivenusianrd_clip_image010.jpg" alt="" width="268" height="268"/></p><br />
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<p>(E) inv-llo strain, merged image of confocal microscope and bright field, 1000X</p></td><br />
<td width="296" valign="top"><p><img src="https://static.igem.org/mediawiki/igem.org/2/2a/Ymivenusianrd_clip_image014.jpg" alt="" width="266" height="266" /> <br /><br />
(F) inv-llo strain, image of confocal microscope, 1000X </p></td><br />
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<td width="318" valign="top"><p><img src="https://static.igem.org/mediawiki/igem.org/a/a0/Ymivenusianrd_clip_image016.jpg" alt="" width="271" height="271" /><br /><br />
(G) inv-llo strain, image of bright field, 1000X</p></td><br />
<td width="296" valign="top"><p><img width="282" height="254" src="https://static.igem.org/mediawiki/igem.org/0/04/Ymivenusianrd_clip_image018.jpg" /> <br /><br />
(H) inv-llo strain, 3D image under confocal microscope, 1000X</p></td><br />
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<tr><br />
<td width="614" colspan="2" valign="top"><p>Figure 3. Red pots were S. elongates PCC7942 under fluorescence microscope. Transparent one was non-EB iPS cells. Both wildtype and inv-llo type could not enter into cells but inv-llo type could trigger itself recruitment to cells.</p></td><br />
</tr><br />
</table><br />
<p>Part 3<br /><br />
</p><br />
<p>Figure 4 revealed the coculture results of S.elongatus PCC 7942 (wt/inv-llo strain)and iPS cells at EB stage.</p><br />
<table border="1" cellspacing="0" cellpadding="0" width="617"><br />
<tr><br />
<td width="304" valign="top"><p><img width="290" height="218" src="https://static.igem.org/mediawiki/igem.org/d/d1/Ymivenusianrd_clip_image002_0001.jpg<br />
" alt="描述: Tv1" /> <br /><br />
(A) iPS cells at EB stage, image of bright field</p></td><br />
<td width="312" valign="top"><p><img width="298" height="224" src="https://static.igem.org/mediawiki/igem.org/4/45/Ymivenusianrd_clip_image004_0001.jpg" /> <br /><br />
(B)inv-llo strain, image of fluorescence</p></td><br />
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<tr><br />
<td width="304" valign="top"><p><img width="290" height="217" src="https://static.igem.org/mediawiki/igem.org/0/0e/Ymivenusianrd_clip_image006_0001.jpg" /> <br /><br />
(C)inv-llo strain, merged image of fluorescence and bright field</p></td><br />
<td width="312" valign="top"><p>Figure 4.<br /><br />
Red pots were S. elongates PCC7942 under fluorescence microscope. Gray one was iPS cells at the EB stage. Results showed that not many inv-llo PCC 7942 could enter into cells. </p></td><br />
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</table><br />
<p>There were similar and relative results of the iPS cells at EB stage and non-EB iPS cells. Both inv-llo type of S.elongatus PCC7942 didn&rsquo;t enter into stem cells but triggered the recruitment to cells. Because embryoid body contained not only one cell, the entry for cyanobacteria would be more difficult but the recruitment provided a potential for invading mammalian cells. Future study will focus on different coculturing condition and after invasion into stem cells, differentiation could render Venusian.</p><br />
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<h2 class="drawer-handle open">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
</ul> <br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
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<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html
Team:NYMU-Taipei/ymivenusianmm.html
2012-10-27T01:51:28Z
<p>Jayms: </p>
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<p><span class="subtitle">What&rsquo;s the specialty of our cyanobacteria?</span></p><br />
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<p>Invasin(protein names: P11922&nbsp; INVA_YERPS) is a protein that allows enteric bacteria to penetrate cultured mammalian cells. Listeriolysin O (LLO) facilitates bacterial escape from the internalization vesicle into the cytoplasm, where bacteria divide and undergo cell-to-cell spreading via actin-based motility[2]. And Synechococcus elongatus PCC 7942 was the first cyanobacterial strain which could be transformed by exogenously added DNA. When engineered with invasin(inv) from&nbsp;<em>Yersinia</em><em>pestis</em>&nbsp;and listeriolysin O(llo) from&nbsp;<em>Listeria monocytogenes (both provided by Pamela A. Silver, Harvard Medical school</em><em>[3]</em><em>)</em>,&nbsp;new <em>S. elongatus</em>&nbsp;was able to invade cultured mammalian cells and was capable of symbiosis with eukaryotic cells. <br /><br />
Additionally, invasin(inv) from&nbsp;<em>Yersinia</em><em>pestis</em>&nbsp;and listeriolysin O(llo) from&nbsp;<em>Listeria monocytogenes </em><em>have been studied by 2010 Warsaw igem team. Our further study and novel observation relative to their research were posted not only on our wiki, but also on the partregistry website(</em><em><u>BBa_K299810</u></em><em>,</em> <u>BBa_K299811</u>,<u>BBa_K299812</u><em>) </em></p><br />
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<p>Three types of mammalian cells were chosen—two stages of induced pluripotent stem cells from mice and J774 mouse macrophage cell line. Here are the protocols of cell culturing (two stages of mice induced pluripotent stem cells: iPS cells separated from embryonic fibroblasts (MEFs) <u>click here,</u> embryoid body (EB) stage of iPS cells <u>click here</u>, J774 mouse macrophage cell line <u>click here</u>)<br /><br />
</p><br />
<p>Cells were seeded on the 24-well plate. After 80% full in each well, PCC7942(wt/inv-llo transform) suspensions in PBS were set to the same OD and 50 µl of this suspension were added per 1 ml of Leibovitz's L-15 medium without phenol red, without antibiotics and with 10% fetal bovine serum (Invitrogen) per well of 24-well tissue culture dishes containing the mammalian cells. After 24-hour cells and PCC7942 coculturing under 30°C atmospheric CO2 level, supernatants were discarded and cells were washed with PBS two times and the medium was replaced by L-15 containing antibiotics, gentamycin 100ug/ml(Invitrogen). After 24 hours, cells were fixed by 4% paraformaldehyde and were observed by fluorescent microscope or confocal laser microscope. </p><br />
<p><img src="https://static.igem.org/mediawiki/2012/f/f5/Ymiv2.png" alt="" width="305" height="485" /><img src="https://static.igem.org/mediawiki/2012/d/d9/Ymiv3.png" alt="" width="321" height="441" /></p><br />
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<h2 class="drawer-handle open">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
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<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
</li><br />
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<br />
<li class="drawer"><br />
<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html
Team:NYMU-Taipei/ymivenusianintro.html
2012-10-27T01:51:17Z
<p>Jayms: </p>
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href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
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<div class="imgbox" style="width:344px;float:right;"> <img src="https://static.igem.org/mediawiki/2012/0/0d/Ymiv1.png" alt="" width="344" height="282" /></div><br />
<p>Cyanobacteria obtain their energy by photosynthesis. In previous research in our study, we hope to create new type Synechococcus elongatus PCC7942 which could do both the nitrogen and sulfide metabolism. This new cyanobacteria would survive on the SO2-rich environment on the Venus. Meanwhile,when engineered with invasin(inv) from <em>Yersinia</em><em>pestis</em> and listeriolysin O(llo) from <em>Listeria monocytogenes (</em><em>both provided by Pamela A. Silver, Harvard Medical school</em><em>)</em>, <em>S. elongates</em> was able to invade cultured mammalian cells.<br /><br />
</p><br />
<p>J774 mouse macrophage cell line, a kind of differentiate cells, could engulf cyanobacteria not only by phagocytosis but speedy by the inv-llo engineering help. Additionally, stem cells can divid and differentiate into diverse specialized cell types and can self-renew to produce more stem cells.&nbsp;&nbsp;In a developing embryo, stem cells can differentiate into all the specialized cells (called pluripotent cells). Induced pluripotent stem cells (iPS cells ) are stem cell-like cells derived from reprogrammed somatic cells. IPS cells are then seeded on mouse embryonic fibroblast (MEF) feeder cells and cultured in ES cell medium. After separation iPS cells from MEF, non-EB iPS cells can be gotten. After culturing non-EB iPS cells on the petri dish, iPS cells at Embryoid Body(EB) stage can be obtained. Embryoid Body (EB) culture is used to examine the differentiation potential of the embryonic stem (ES) cells.<br /><br />
</p><br />
<p>Thus, if engineered cyanobacteria can invade into stem cells and produce energy and O2 by photosynthesis, a dream of creating Venusians would not be far.</p><br />
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<h2 class="drawer-handle open">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
</ul> <br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
</li><br />
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<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
</ul><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html
Team:NYMU-Taipei/ymivenusian.html
2012-10-27T01:51:07Z
<p>Jayms: </p>
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<p>Are there <em><u>Venusian</u></em>? Is it possible that there are creatures living on the Venus? Dreams will come true by the symbiosis of cyanobacteria and eukaryotes.<br /><br />
</p><br />
<p>Cyanobacteria obtain their energy by photosynthesis. Sulfide-dependent photosynthesis cyanobacteria could resist the harmful environment and do the electron transport by sulfide quinine reductase(SQR), which we cloned previously. Additionally, references revealedwhen engineered with invasin(inv) from <em>Yersinia</em><em>pestis</em> and listeriolysin O(llo) from <em>Listeria monocytogenes</em>, <em>S. elongates</em> was able to invade cultured mammalian cells.<br /><br />
</p><br />
<p>The symbiosis possibility of induced pluripotent stem cells from mice and J774 mouse macrophage cell line were evaluated. Results showed that invasin(<em>inv</em>,from <em>Yersinia</em><em>pestis</em>) and listeriolysin O(llo, from <em>Listeria monocytogenes</em>) transformation would raise the entry speed and escape rate in J774 mouse macrophage cell line, and would increase the recruitment of cyanobacteria on induced pluripotent stem cells. </p><br />
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<h2 class="drawer-handle open">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
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<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Cd+2 Collector</h2><br />
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<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymic5.html
Team:NYMU-Taipei/ymic5.html
2012-10-27T01:50:56Z
<p>Jayms: </p>
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href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
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<ol><br />
<li>We have successfully cloned zinTp into pSB1C3</li><br />
<li>We have successfully cloned zinTp-GFP into pSB1C3</li><br />
<li>We have characterized zinTp function. The expression of downstream gene will increase when added cadmium ion into LB medium, and the maximum transcription rate occurs around 60 min after added 200μM cadmium.</li><br />
<li>We have successfully cloned mnt into pSB1C3</li><br />
<li>We have successfully cloned smtA into pSB1C3</li><br />
<li>We have characterized smtA function. When expressing smtA, it enhances E.coli tolerance to cadmium.<br /><br />
</ol> <br /><br />
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<span class="subtitle">References</span></div><br />
<br /><br />
Sigel, A.; Sigel, H.; Sigel, R.K.O., ed. (2009). <em>Metallothioneins and Related Chelators</em>. Metal Ions in Life Sciences. Cambridge: RSC Publishing<br /><br />
<br /><br />
Jianguo Shi, William P. Lindsay, James W. Huckle, Andrew P. Morby and Nigel J. Robinson. Cyanobacterial metallothionein gene expressed in Escherichia coli Metal-binding properties of the expressed protein. FEB.5 11081 Volume 303, number 2,3, 159-163<br /><br />
<br /><br />
Sode et al. (1998) Construction of a marine cyanobacterial strain with increased heavy metal ion tolerance by introducing exogenic metallothionein gene. <em>J Mar Biotechnol</em> <br /><br />
<br /><br />
ZHIQI HAO,1 SHAOLIN CHEN,1 AND DAVID B. WILSON1,2, Cloning, Expression, and Characterization of Cadmium and Manganese Uptake Genes from <em>Lactobacillus plantarum,</em> APPLIED AND ENVIRONMENTAL MICROBIOLOGY,Nov. 1999, p.4746–4752 Vol. 65, No. 11 <br /><br />
<br /><br />
A. Paskarova, P. Ferianc, J. Kormanec, D. Homerova, A. Farewell, T. Nystrom. Regulation of yodA encoding a novel cadmium-induced protein in Escherichia coli, Microbiology (2002), 148, 3801–3811 <br /><br />
<br /><br />
Damien Arnoult, Ire`ne Tatischeff, Je´rome Estaquier, Mathilde Girard, Franck Sureau, Jean Pierre Tissier, Alain Grodet, Marc Dellinger, Francois Traincard,Axel Kahn,Jean-Claude Ameisen, and Patrice Xavier Petit. &ldquo;On the Evolutionary Conservation of the Cell Death Pathway: Mitochondrial Release of an Apoptosis-inducing Factor during <em>Dictyostelium discoideum</em> Cell Death&rdquo;, Molecular Biology of the Cell Vol. 12, 3016–3030, October 2001<br /> <br /><br />
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<h2 class="drawer-handle open">Cd+2 Collector</h2><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymic4.html
Team:NYMU-Taipei/ymic4.html
2012-10-27T01:50:47Z
<p>Jayms: </p>
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<div class="title">Results & Discussion</div><br />
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<p><span class="subtitle">Cloning zinTp into pSB1C3 plasmid</span></p><br />
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<div align="left"><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/a/a3/Ymic13.png" width="600" height="220" /><br /><br />
We got the right plasmid which contained zinTp<br /><br />
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<div class=out style='text-align:center'></div><br />
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<p><span class="subtitle">Characterizing of zinTp</span></p><br />
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<div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/3/3c/Ymic14.png" width="600" height="306" /><br /><br />
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<div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/5/53/Ymic15.png" alt="" width="494" height="348" /><br /><br />
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<p>In this fig we could see that before 60 min, the fluorescence level of zinTp+GFP steadily increased, while BBa_I13522 (positive control) remained stable around 35000 au as time gone by. At 60 min, zinTp reached its maximum transcription rate so the fluorescence level stop increasing around 55000 au.</p><div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/a/a0/Ymic16.png" alt="" width="494" height="306" /><br /><br />
<strong>Fluorescence level at 15mins after Cd2+ added </strong><br /><br />
<br />
</div><br />
<p>This fig shows the fluorescence level of different cadmium concentration. We can see that BBa_I13522 remained stable from different cadmium concentration, while the fluorescence level of zinTp+GFP increased as the cadmium concentration increased. We could also tell that the transcription rate of zinTp is fairly low when there is no cadmium ion. <br /><br />
<br /><br />
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<p><span class="subtitle">Cloning mnt into pSB1C3 plasmid</span><br /><br />
</p><br />
</div><div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/c/c2/Ymic17.png" width="630" height="243" /><br /><br />
Mnt gene was successfully cloned into pSB1C3 plasmid<br /><br />
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<p><span class="subtitle">Cloning mnt into pSB1C3 plasmid</span><br /><br />
</p><br />
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<div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/c/c3/Ymic18.png" width="600" height="331" /><br /><br />
smtA gene was successfully cloned into pSB1C3 plasmid<br /><br />
</div><br />
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<p><span class="subtitle">Characterizing of smtA</span><br /><br />
</p><br />
</div><div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/2/25/Ymic19.png" alt="" width="508" height="342" /><br /><br />
<strong>Cadmium 0/200/500μM Medium</strong><br /><br />
</div><br />
<br /><br />
<p>This fig shows that smtA enhances the tolerance level to cadmium ion. In wild type E.coli, OD595 differ from between 0/200/500μM cadmium concentration. E.coli hardly grow in 500μM cadmium medium. However, the E.coli which can express smtA seem to show no significant different between 0 &amp; 200μM cadmium medium.</p><br />
<div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/2/26/Ymic20.png" alt="" width="463" height="282" /><br /><br />
<strong>8 hours after cadmium added</strong><br /><br />
</div><br />
<p>OD of WT E.coli decrease when cadmium concentration increase. The E.coli which can express smtA seem to have stronger resistance to cadmium toxicity and the E.coli concentration show no significant different from 0 to 200μM cadmium medium.</p><br />
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<div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/1/14/Ymic21.png" alt="" width="386" height="291" /><br /><br />
<em>E.coli</em> which could express smtA growth better than wild type in 200μM Cd+2 medium <br /><br />
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<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
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<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
</li><br />
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<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
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<li class="drawer last"><br />
<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymic3.html
Team:NYMU-Taipei/ymic3.html
2012-10-27T01:50:37Z
<p>Jayms: </p>
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<span class="subtitle">Kill-switch of amoeba</span><br /><br />
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<p><strong>Cloning zinTp into psb1c3 plasmid</strong><br /><br />
We got the zinTp sequence from RegulonDB <a title="http://ppt.cc/SS5N" >(1)</a>. Amplification was performed for 40 cycles of denaturation for 30 sec at 94 °C, annealing for 1 min at 59 °C and polymerization for 1 min at 68 °C. A PCR product containing the zinTp was cloned into plasmid psb1c3. The plasmid we used was got from iGEM 2012 distribution kit. The resulting plasmid was transformed into E. coli DH5αcompetent cells.<br />
<br /><br />
<br /><br />
<strong>Characterizing of zinTp</strong><br /><br />
We ligated the zinTp with GFP generator(BBa_E0840) and tested the function of zinTp. The transgenic E.coli contains zinTp+GFP, which should fluoresce when Cd2+ exist. We added different concentration of cadmium ion into LB medium and compared the fluorescence level of E.coli which contain zinTp+GFP, pTetR+GFP (BBa_I13522), and psb1c3 backbone only(wildtype control). </p><br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/f/f0/Ymic10.png" alt="" width="495" height="311" /><br /><br />
<p align="left">We grew E.coli to OD595=0.3~0.4, then added cadmium ion into micro centrifuge tubes. The E.coli was incubated at 37°C. We tested the fluorescence level of GFP. The excitation wavelength is 501nm and the emission wavelength is 511nm. <br /><br />
<br /><br />
<br /><br />
</p><br />
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<div align="left"><br />
<span class="subtitle">Enhancing absorption and Tolerance to cadmium ions</span><br /><br />
<br /></div><br />
<p><strong>Cloning mnt into psb1c3 plasmid</strong><br /><br />
We got the mnt sequence from NCBI<a title="http://ppt.cc/Unhg" >(2)</a><a title="http://ppt.cc/feOU" >(3)</a><a title="http://ppt.cc/9-X5" >(4).</a> Next, we grew <em>Synechocystis sp. PCC 6803</em> and cloned the mnt gene into psb1c3. Then we transformed the plasmid into E. coli DH5αcompetent cells. The plasmid we used was got from iGEM 2012 distribution kit.<br /><br />
<br /><br />
<strong></strong><strong>Cloning smtA into psb1c3 plasmid<br /><br />
</strong>Firstly, we got the smtA sequence from NCBI <a title="http://ppt.cc/t9kR" >(5)</a>. Next, we grew <em>Synechococcus elongatus</em> PCC 7942 and cloned the smtA gene into psb1c3. Then we transformed the plasmid into E. coli DH5αcompetent cells. <br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/f/ff/Ymic11.png" alt="" width="558" height="357" /><br /><br />
</p><br />
</div><br />
<strong>Characterizing of smtA<br /><br />
</strong>We cloned the smtA gene into the plasmid Ptrc-PNSII-Strep. This plasmid includes a promoter Ptrc. We compared the tolerance level between Ptrc-smtA-PNSII-Strep E.coli and E.coli that did not contain smtA(Ptrc-PNSII-Strep plasmid as a wild type control). We tested the growth curve of E.coli with different cadmium concentration.<br /><br />
<br /><div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/4/4e/Ymic12.png" alt="" width="395" height="492" /></p><br />
<p align="left">The E.coli was grown to OD595=0.3~0.4, then we added 200λ broth into 6.cc liquid LB medium with Streptomycin 25 μg/ml. We incubated the E.coli at 37 °C.</p><br />
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<h2 class="drawer-handle open">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
</li><br />
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<li class="drawer last"><br />
<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymic2.html
Team:NYMU-Taipei/ymic2.html
2012-10-27T01:50:22Z
<p>Jayms: </p>
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<p><strong><em>Into the cell<br />
</em></strong><br /><br />
By cloning LLO and invasin into the E.coli, the bacteria can get into the amoeba and live inside it. Invasin helps the cell to be endocytosed by the host. Then Listeriolysin O makes the cell escaped from phagosomes in the cytoplasm of the host. <br /><br />
<br /><br />
<strong><em>Cadmium accumulation</em></strong><br /><br />
To enhanced E.coli&rsquo;s ability of gathering cadmium ions, we clone smtA and mnt gene into the E.coli. SmtA is a cadmium binding metallothionein, which can bind to cadmium ions. Mnt is an ion transporter that pumps cadmium ions inside the bacteria.</p><br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/6/6b/Ymic3.png" alt="" width="471" height="292" /><br /><br />
The main idea of how E.coli and Dictyostelium discoideum system works.<br /><br />
<br /><br />
</p><br />
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<p> <br /><br />
---<strong><em>smtA</em></strong>---<br /><br />
<strong></strong>Metallothionein (MT) is a family of cysteine-rich, low molecular weight (MW ranging from 500 to 14000 Da) proteins. MTs have the capacity to bind both physiological (such as zinc, copper, selenium) and xenobiotic (such as cadmium, mercury, silver, arsenic) heavy metals through the thiol group of its cysteine residues<a title="Sigel, A.; Sigel, H.; Sigel, R.K.O., ed. (2009). Metallothioneins and Related Chelators. Metal Ions in Life Sciences. 5. Cambridge: RSC Publishing." >(1)</a>.<br /><br />
Synechococcus PCC. 7942 gene smtA encodes the protein designated to be a metallothionein (MT). While expressing this gene in E.coli, it can increase cadmium ion tolerance of E.coli and so does the accumulation of cadmium ion<a title="Jianguo Shi, William P. Lindsay, James W. Huckle, Andrew P. Morby and Nigel J. Robinson. Cyanobacterial metallothionein gene expressed in Escherichia coli Metal-binding properties of the expressed protein. FEB.5 11081 Volume 303, number 2,3, 159-163" >(2)</a><a title="Sode et al. (1998) Construction of a marine cyanobacterial strain with increased heavy metal ion tolerance by introducing exogenic metallothionein gene. J Mar Biotechnol" >(3)</a>.<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/1/1f/Ymic4.png" alt="" width="359" height="223" /><br /><br />
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<br /><br />
---<strong>mnt</strong>---<br /><br />
Mnt gene is the Mg2+/Cd2+ transporter which belongs to ABC transporters system. The ATP-binding cassette (ABC) transporters form one of the largest known protein families, and are widespread in bacteria, archaea, and eukaryotes. They couple ATP hydrolysis to active transport. The structure of a prokaryotic ABC transporter usually consists of three components; typically two integral membrane proteins each having six transmembrane segments, two peripheral proteins that bind and hydrolyze ATP, and a periplasmic (or lipoprotein) substrate-binding protein. Many of the genes for the three components form operons as in fact observed in many bacterial and archaeal genomes.<a title="http://ppt.cc/kBfk" >(4)</a><br /><br />
<br /><div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/1/1f/Ymic5.png" alt="" width="325" height="362" /></p><br />
<p>Cloning mntA gene into E.coli can increase Cd2+ uptake. </p><br />
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<span class="subtitle">Biosafety Consideration</span></div><br />
<p><strong>---</strong> <strong>Precaution against </strong><strong>Horizontal gene transfer – Living inside of amoeba</strong>---<br /><br />
Horizontal gene transfer(HGT) is a serious issue concerning the release of genetically modified organisms (GMOs) into the environment. To prevent HGT, we must make sure our genetic modified E.coli will never join their wild type relations. Our solution is to put the engineered E.coli into amoeba, so the E.coli cannot contact the environment directly. Whenever wild type bacteria break into the amoeba, they will be digested.<br /><div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/7/76/Ymic6.png" alt="" width="382" height="246" /></p><br />
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<p><strong><br /><br />
--- Precaution against becoming invasive species - Kill switch of Amoeba</strong>---<br /><br />
To make sure our genetic modified organisms will stay in the area where we want them to clean up cadmium ions, we also build a kill-switch that will switch on when there is no cadmium ion in the soil. The kill-switch has two man-made operons. The first one includes zinTp(previous name pYodA), this promoter activity is induce by cadmium ion; and its constructive gene is lacI. The other one includes BBa_R0010, the promoter whose activity is inhibited by the protein lacI; and the constructive gene DdaifA and HlyA(BBa_K223054). DdaifA is a protein that cause the apoptosis of Dictyostelium discoideum cell. HlyA is a tag that stick after the target protein. With this tag, E.coli will secrete the target protein. When cadmium ions exist, zinTp works and lacI is generated, so the expression of aifA and HlyA is repressed. If there is no cadmium ions, BBa_R0010 will activate the expression of aifA and HlyA, so the protein aifA will be secrete right into the cytoplasm of amoeba, causing amoeba apoptosis.<br /><br />
</p><br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/4/47/Ymic7.png" width="500" height="385" /></p><br />
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<p> <br /><br />
---<strong><em>ZinTp</em></strong>---<br /></p><br />
<p>ZinTp (pYodA) is a promoter which expresses the downstream gene in the presence of cadmium ion. The activity of this promoter is specific affect by cadmium ion and won&rsquo;t be induced by other ions like zinc, copper, cobalt, and nickel<a title="A. Paskarova, P. Ferianc, J. Kormanec, D. Homerova, A. Farewell, T. Nystrom. Regulation of yodA encoding a novel cadmium-induced protein in Escherichia coli, Microbiology (2002), 148, 3801–3811" >(5)</a>. <div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/5/5f/Ymic80.png" alt="" width="300" height="300" /></p><br />
<p align="left">Strain AL6 (λФ PyodA–lacZ) was grown aerobically in LB medium at 37°C in presence (●) or absence (○) of cadmium (136±5μM) <em>β</em>-Galactosidase activity in the<br />
presence (■) or absence (□) of tested agents was also measured.</p><br />
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<p> <br /><br />
---<strong><em>DdaifA</em></strong>---<br /></p><br />
<p>Apoptosis-inducing factor (AIF) is involved in a caspase-independent cell death pathway. <em>Dictyostelium discoideum</em> has a homolog of mammalian AIF, which could cause nuclei breakdown, leading the cell to the apoptosis program<a title="Damien Arnoult, Ire`ne Tatischeff, Je´rome Estaquier, Mathilde Girard, Franck Sureau, Jean Pierre Tissier, Alain Grodet, Marc Dellinger, Francois Traincard,Axel Kahn,Jean-Claude Ameisen, and Patrice Xavier Petit. “On the Evolutionary Conservation of the Cell Death Pathway: Mitochondrial Release of an Apoptosis-inducing Factor during Dictyostelium discoideum Cell Death”, Molecular Biology of the Cell Vol. 12, 3016–3030, October 2001" >(6)</a>.<br /><div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/f/ff/Ymic9.png" alt="" width="202" height="225" /></p><br />
<p align="left">DdAIF induces damage of mammalian and Dictyostelium nuclei in a cell-free system. (E) shows the quantification of damaged nuclei induced by cytoplasmic extracts.<br />
Damaged nuclei were assessed by the disappearance of the nucleoli. One hundred nuclei were counted in each condition. Histograms are the mean of three independent experiments. Nu, nucleolus; Dnu, dying cells nucleolus. Dictyostelium cells (control CE), Dictyostelium cells treated with PPIX to cause cell death (dying cells CE). Cytoplasmic extracts of dying cells were also immunodepleted with anti-DdAIF (dying cells CE -AIF) or with nonimmune serum (dying cells CE NIS)<br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymic1.html
Team:NYMU-Taipei/ymic1.html
2012-10-27T01:50:14Z
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<p>We&rsquo;ve built the endosymbiotic system to clean up cadmium contamination. The engineered organism can collect cadmium ion in the soil and can be easily gather together; moreover, the device won&rsquo;t become another pollution source to our environment.<br /><br />
Cadmium contamination is a serious problem in many countries; cadmium can cause respiratory system damage, renal failure, bones softening, and, maybe the most infamous case in Asia, Itai-itai disease. The rice absorbed the cadmium in the soil and the cadmium accumulated in the people eating contaminated rice. The aim of our project is to build up a system that can collect Cadmium ion in the soil and can be easily gather together; besides, we don&rsquo;t want our device become another pollution source to our environment. We found that the endosymbiotic system between the soil-living amoeba <em>Dictyostelium discoideum </em>and <em>E.coli </em>would be our favorite solution.<br /><br />
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Cadmium ions were absorbed by amoeba then accumulated inside of E.coli<br /><br />
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<p><span class="subtitle">Why <em>Dictyostelium discoideum</em></span></p><br />
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<em>Dictyostelium discoideum </em>is a soil-living amoeba, so it is a good choice to collect cadmium ion in soil. In addition, the amoeba naturally has endosymbiont inside the cell. The E.coli living in amoeba accumulates cadmium ions, just like an organelle, the E.coli acts as a compartment preventing its host from cadmium poisoning. Furthermore, keeping the E.coli inside the amoeba avoid unexpectedly gene exchange between engineered E.coli and other wild types bacteria. Moreover, Dictyostelium discoideum chemotaxis is well-studied and can be easily gather by adding cAMP.<br /><br />
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<p>Single cell amoeba <em>Dictyostelium discoideum</em> gathered when cAMP was added.<br /><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymin4.html
Team:NYMU-Taipei/ymin4.html
2012-10-27T01:50:03Z
<p>Jayms: </p>
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<a <br />
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<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymim1" target="_parent">Modeling</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymihpa.html">Human Practice</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
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<p><span class="subtitle">Practical Application</span></p><br />
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<p>Moreover, our project has been put into practice thanks to the cooperation with Chung Hwa Pulp Corporation. The wastewater generated by Pulp factories contains enormous sulfide compounds and nitrate, which bring the annoying odors as well as the contamination to the local environment. Fortunately, our project seems to be the solution to their problem. This also demonstrates the potential and possibility of commercialized our project. On top of that, the engineered cyanobacteria can become the third endosymbiosis organelles with the help of division inhibitor, gene for invasion. After installing our designation into plants or even human cells as artificial organelles, we grant eukaryotes the ability to survive in extreme environments as horrible as Venus in case the space immigration is necessary on day.<br /><br />
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<p><span class="subtitle">References</span></p><br />
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<p><em>Sulfur-driven autotrophic denitrification: diversity ,biochemistry, and engineering applications</em> (1 September 2010) by Ming-Fei Shao &amp; Tong Zhang &amp; Herbert Han-Ping Fang <br />
<p><em>Characterization of the membranous denitrification enzymes nitritereductase (cytochrome cd1) and copper-containing nitrous oxidereductase from Thiobacillus denitrificans </em>(30 October 1995) by Ursula H. Hole · Kai-Uwe Vollack · Walter G. Zumft ·Effi Eisenmann · Roman A. Siddiqui ·Bärbel Friedrich · Peter M. H. Kroneck</p><br />
<p><em>Autotrophic denitrification for combined hydrogen sulfideremoval from biogas and post-denitrification </em>by R. Kleerebezem and R. Mendez (2002)</p><br />
<p><em>Nitrous oxide production and consumption: regulation of gene expression by gas-sensitive transcription factors </em>(7 July 2012) by Stephen Spiro</p><br />
<p><em>Mechanism of Sulfide-Quinone Reductase Investigated Using Site-Directed Mutagenesis and Sulfur Analysis</em> (6 August 2002)by Christoph Griesbeck, Michael Schu¨tz, Thomas Scho¨dl, Stephan Bathe, Lydia Nausch, Nicola Mederer, Martin Vielreicher, and Gu¨nter Hauska</p></div><br />
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<h2 class="drawer-handle open">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
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<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
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<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymin3.html
Team:NYMU-Taipei/ymin3.html
2012-10-27T01:49:49Z
<p>Jayms: </p>
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<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymim1" target="_parent">Modeling</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymihpa.html">Human Practice</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
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<p><span class="subtitle">Part : BBa_K896004</span></p><br />
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<p>NosZ-NorCB<br /><br />
In<em> Pseudomonas aeruginosa PAO1</em>, NorCB is found as two sub-units – nitric oxide reductase subunit C and B. As a matter of fact, these two subunits are adjacent genes. And thus we cloned them together to produce a functional reductase. <div class=out style='text-align:center'><br />
<img border="0" src="https://static.igem.org/mediawiki/2012/9/9d/Ymin4.gif" align="center" alt="" /><br /><br />
http://www.stanford.edu/group/collman/nor.htm<br />
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<p>NosZ is the structure gene of nitrous reductase. Very few microorganisms process similar enzyme, and many of them are homologs according to NCBI database. Furthermore, it performs the last procedure of bio-denitrification – nitrous oxides to nitrogen – in our model. <br />
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Obviously, the reason we cloned NorCB and NosZ on the same plasmid is to connect the continuous denitrifying procedures together.</p><br />
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<img src="https://static.igem.org/mediawiki/2012/5/52/Ymin5.gif" alt="" /><br /><br />
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<p><a href="http://partsregistry.org/Part:BBa_K896004">http://partsregistry.org/Part:BBa_K896004</a><br /><br />
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<p><span class="subtitle">Part:BBa_K896006</span></p><br />
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<p>NirN-NirS<br /><br />
Nitrite reductase is composed of several different subunits. Together, it can reduce nitrite to nitric oxide. We clone all these subunits together on the same plasmid in order to grant cyanobacteria a functional protein.</p><br />
<div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/4/42/Ymin6.gif" alt="" width="500" border="0" align="center" /><a href="http://montypython.scs.uiuc.edu/Publications/pubs.php">http://montypython.scs.uiuc.edu/Publications/pubs.php</a><br /><br />
<a href="http://partsregistry.org/Part:BBa_K896006">http://partsregistry.org/Part:BBa_K896006</a><br /><br />
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<p><span class="subtitle">Part:BBa_K896007</span></p><br />
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<p>Nap( Nitrate->Nitrite reductase)<br /><br />
Nap is periplasmic nitrate reductase, which is known for the ability to reduce nitrate into nitrite. In microorganisms, similar reductases are often observed. For instance, even E. coli has its own nitrate reductase and can use nitrate as final electron acceptor. Moreover, we acquire the Nap gene from Desulfovibrio desulfuricans ATCC 27774.</p><br />
<div class="out" style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/d/d9/Ymin7.gif" alt="" width="399" border="0" align="center" /><br /><br />
<p><a href="http://partsregistry.org/Part:BBa_K896007">http://partsregistry.org/Part:BBa_K896007</a><br /><br />
</p></div><br />
Since Nir and Nap complex together would link the reducing reaction from Nitrate to nitric oxide, we ligated these two gene together and expressed them on <em>Synechococcus elongatus PCC 7942</em>.</div><br />
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<h2 class="drawer-handle open">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymin2.html
Team:NYMU-Taipei/ymin2.html
2012-10-27T01:49:40Z
<p>Jayms: </p>
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<a <br />
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<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymim1" target="_parent">Modeling</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymihpa.html">Human Practice</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
<a href="https://2012.igem.org/Main_Page">iGEM</a><br />
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<div class="title">Methods</div><br />
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<p>Recently, the search for biological nitrogen removal method from wastewaters and exhaust air has come up with several promising methods; however, most of them just took advantage of some special bacteria combined with industrial procedures. On the contrary, our iGEM project aim to reduce nitrogen oxides and oxidize sulfide compounds at the same time. <br /><br />
<br /><br />
During the processes of denitrification, sulfide compounds and nitrate act as electron donor and acceptor respectively. This reaction is also known as sulfide-driven denitrification. Researchers have reported that E. coli can perform such reaction when expresses sqr gene from <em>R. capsulatus</em>. Herein, we enable certain type of cyanobacteria to take advantage of sulfide and reduce nitrogen oxide compounds into nitrogen. The BLAST result shows that sqr genes are homolog in <em>R. capsulatus</em> and <em>Synechocystis sp. PCC 6803</em>. <br />
For denitrification, we plan to get access to <em>Thiobacillus denitrificans</em>, the well-known chemolithotrophic organisms. Nevertheless, we later found it difficult to obtain the specific strain we need. According to NCBI database, enzymes for denitrification such as nir, nor, nos share great similarity between <em>Thiobacillus denitrificans </em>and <em>Pseudomonas aeruginosa PAO1</em>, so we adopted <em>P. aeruginosa PAO1</em> instead and expressed the enzymes mentioned above in <em>Synechocystis sp. PCC 6803</em>.<br /><br />
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<div class=out style='text-align:center'><br />
<img src="https://static.igem.org/mediawiki/igem.org/6/6c/Ymin2.png" alt="" width="554" height="190" border="0" align="center" /><br /><br />
<p><a href="http://www.genome.jp/kegg/pathway/map/map00910.html">http://www.genome.jp/kegg/pathway/map/map00910.html</a><br /><br />
</p><br />
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<p><br /><br />
These denitrifying enzymes are functional under aerobic condition, yet like all cyanobacteria, <em>Synechocystis sp. PCC 6803</em> produces oxygen during photosynthesis. Fortunately, when sulfide presents in the environment and sqr is expressed, it will cease producing oxygen and use sulfide as an electron donor for carbon dioxide photoassimilation. Together with dsrI and dsrII enzymes from <em>Desulfovibrio desulfuricans</em>, our engineered organisms are capable of reducing three major oxides pollution – nitrogen, sulfur and carbon oxides. <br /><br />
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<p><img src="https://static.igem.org/mediawiki/igem.org/a/ab/Ymin3.png" alt="" border="0" align="center" /><br /><br />
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<p><a href="http://fleasnobbery.blogspot.tw/2008/07/cyanobacteria.html">http://fleasnobbery.blogspot.tw/2008/07/cyanobacteria.html</a></p><br />
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<h2 class="drawer-handle open">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymin1.html
Team:NYMU-Taipei/ymin1.html
2012-10-27T01:49:31Z
<p>Jayms: </p>
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<head><br />
<meta http-equiv="Content-Type" content="text/html; charset=utf-8" /><br />
<title>NYMU iGEM</title><br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymim1" target="_parent">Modeling</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymihpa.html">Human Practice</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
<a href="https://2012.igem.org/Main_Page">iGEM</a><br />
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<div class="title">Background</div><br />
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<p>Nitrogen oxides are one of the most notorious pollutants in the modern days. Anthropogenic disturbance of Nitrogen cycle accelerates the accumulation of nitrogen oxides, and such situation will deteriorate further if we aren’t devoted to solve the environmental challenge. <div class=out style='text-align:center'><br />
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<h2 class="drawer-handle open">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
</ul> <br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymiq4.html
Team:NYMU-Taipei/ymiq4.html
2012-10-27T01:49:09Z
<p>Jayms: </p>
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<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymiproject.html" target="_parent">Project Venusian</a> ·<br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymim1" target="_parent">Modeling</a> · <br />
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href="https://2012.igem.org/Team:NYMU-Taipei/ymihpa.html">Human Practice</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymijf.html">Extras</a> · <br />
<a <br />
href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
<a href="https://2012.igem.org/Main_Page">iGEM</a><br />
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<div class="title">Results &amp; References</div><br />
<div align="left"><br />
<p><span class="subtitle">Sulfide concentration and the growth of sqr expressing strain Synechococcus sp. PCC 7942</span></p><br />
</div><br />
<div align="left"><br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/9/96/Ymiq5-1.png" alt="" width="557" height="299" /><br /><br />
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<p align="center"><img src="https://static.igem.org/mediawiki/2012/1/16/Ymiq5-2.png" alt="" width="557" height="312" /><br /><br />
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<p>The horizontal axis stands for day passed, while the vertical axis is the absorbance of an OD 730nm. According to the graph, sqr expressing strain grew much better than wild type strain when sulfide was added into the medium. This result suggests that sqr is not only expressed but functional in the cyanobacteria.<br /><br />
<br /><br />
<br /><br />
</p></div><br />
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<p><span class="subtitle">Standard curve</span></p><br />
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<p align="center"><img src="https://static.igem.org/mediawiki/2012/8/81/Ymiq6.png" width="626" /><br /><br />
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<p>We established a H2S standard curve to quantify H2S concentration inside our experiments.<br /><br />
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<div align="left"><br />
<p><span class="subtitle"> Lab Incubation with Escherichia coli</span></p><br />
</div><br />
<br />
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(1) Different Concentration under 24HR<br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/f/f3/Ymiq7.png" width="510" /><br /><br />
</p><br />
</div><br />
<p>We used different H2S concentration challenge ptrc-sqr transformed E.coli and tested H2S consumption in 24hrs. The result shows that ptrc-sqr transformed E.coli consumed much more H2S compred to the blank control, ptrc-str transformed E.coli. In other words, we believe our sqr gene works!!<br /><br />
<br /><br />
<br /><br />
(2)<br />
Different Concentration under 48HR<br />
</p><div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/a/ac/Ymiq8-1.png" width="557" height="362" /><br /><br />
</p><br />
</div><br />
<br />
<br />
We followed the experiments in 48hrs and analyzed the H2S consumption in different groups. As the same as our expected, in 48hrs, our ptrc-sqr transformed E.coli depleted much more H2S than 24hrs timepoint.<br /><br />
<br /><br />
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<div align="left"><br />
<p><span class="subtitle"> Differences Between 24 HR and 48 HR under Same Concentration of H2S</span></p><br />
</div><br />
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<div align="left"><br />
(1) 500mM H2S <br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/0/01/Ymiq9.png" width="628" height="153" /><br /><br />
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<p align="center"><img src="https://static.igem.org/mediawiki/2012/f/fa/Ymiq9-1.png" width="557" height="299" /><br /><br />
</p><br />
</div><br />
<p>If we analyze two group in the same H2S concentration, it&rsquo;s easier to find out that our ptrc-sqr transformed E.coli consumed H2S dramatically.<br /><br />
<br /><br />
<br /><br />
(2)<br />
250mM H2S </p><br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/4/49/Ymiq10.png" width="628" height="153" /><br /><br />
</p><br />
</div><br />
<br />
<br />
The experiment result are the same as previous group, ptrc-sqr transformed E.coli can consume much more H2S. <br /><br />
<br /><br />
<br /><br />
(2)<br />
125mM H2S </p><br />
<div class=out style='text-align:center'><br />
<p align="center"><img src="https://static.igem.org/mediawiki/2012/a/a6/Ymiq11.png" width="628" height="177" /><br /><br />
</p><br />
</div><br />
<p>This diagram illustrates that even at a concentration of 125mM of sodium sulfide, the difference of consumption rate is obvious between sqr expressing strain and wild type one.</p><br />
<br /><br />
</div><br />
<br />
<div align="left"><br />
<p><span class="subtitle"> References</span></p><br />
</div><br />
<br />
<br />
<br />
<div align="left"><em>Facultative Anoxygenic Photosynthesis in the Cyanobacterium Oscillatoria limnetica </em>YEHUDA COHEN,* ETANA PADAN, AND MOSHE SHILO Department of Microbiological Chemistry, The Hebrew University-Hadassah Medical School, Jerusalem, Israel Received for publication 9 May 1975<br /><br />
<br /><br />
<em>Sulfide oxidation in gram-negative bacteria by expression of the sulfide–quinone reductase gene of Rhodobacter capsulatus and by electron transport to ubiquinone</em> Hiroomi Shibata and Shigeki Kobayashi 2001<br /><br />
<br /><br />
<em>Sulfur metabolism in Thiorhodoceae. I. Quantitative measurements on growing cells of Chromatium okenii</em>. Antonie Leeuwenhoek, 30: 225–238 Trüper, H.G., and Schlegel, H.G. 1964.<br /><br />
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<h2 class="drawer-handle open">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
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<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
</li><br />
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<br />
<li class="drawer"><br />
<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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</ul><br />
</li><br />
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<li class="drawer last"><br />
<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
</ul> <br />
</li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymiq3.html
Team:NYMU-Taipei/ymiq3.html
2012-10-27T01:48:59Z
<p>Jayms: </p>
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<div class="title">Experiments</div><br />
<div align="left"><br />
<p><span class="subtitle">Resistance of Cyanobacteria (Synechococcus SP. PCC 7002) to Sulfide compound</span></p><br />
</div><br />
<div align="left"><br />
<p>Synechococcus sp. PCC7002 was cultured in the medium A2, a modification of Stevens et al. (1973) on a rotary shaker (100 rpm). The initial concentration of cells is controlled to an OD730 of 0.1 in fresh medium. Different concentration of sodium sulfide is added, and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) was put into experimental group. Cells are grown in the six-well-plates and monitored under OD730 every 24 hour.<br /><br />
<br /><br />
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<div align="left"><br />
<p><span class="subtitle">Resistance of Synechococcus SP. PCC 7002 to 3 - (3,4-dichlorophenyl) - 1,1 - dimethylurea</span></p><br />
</div><br />
<p>Synechococcus sp. PCC7002 was cultured in the medium A2, rotary shaker and initial cell concentration are the same as the last experiment. DCMU is diluted with A2 medium into different concentration, and 10 mM of sodium sulfide is added to the experimental group. The measurement method and frequency of cell growth has been described previously. <br /><br />
<br /><br />
<div align="left"><br />
<p><span class="subtitle">Sodium sulfide concentration and cell growth</span></p><br />
</div><br />
<p>With the result of previous experiment, the concentration of both sodium sulfide and DCMU is adjusted. Sodium sulfide is diluted from 40mM to 2.5mM, while DCMU 0.5 μM is added into the experimental group.<br /><br />
<br /><br />
</p><br />
<div align="left"><br />
<p><span class="subtitle">The effect of sodium sulfide on Synechococcus SP. PCC 7942 growth rate</span></p><br />
</div><br />
<br />
In this experiment, we aimed to examine whether the sqr Synechococcus SP. PCC 7942 expressed is functional. Synechococcus SP. PCC 7942 with sqr and wild type was cultivated in BG-11 medium (Allen M.M. 1968) on the same rotary shaker described in the first experiment. Initial cell concentration is fixed to an OD730 of 0.1 in the medium. 2.5mM of sodium sulfide is added into the experimental group, and DCMU is diluted from 1μM to 0.5μM. <br />
<br />
<br /><br />
<br />
<div align="left"><br />
<p><span class="subtitle">DCMU concentration and cell growth</span></p><br />
</div><br />
<p>From the last experiment, we found it necessary to perform another independent experiment to find the optimal DCMU concentration of Synechococcus SP. PCC 7942. In addition, the initial cell concentration was adjusted to an OD730 of 0.2 in BG-11 medium. DCMU is diluted from 1μM to 0.125μM. Except of cell concentration, all condition remained the same for both sqr and wild type strain of Synechococcus SP. PCC 7942.<br /><br />
<br /><br />
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<div align="left"><br />
<p><span class="subtitle">Sulfide concentration and the growth of sqr expressing strain Synechococcus SP. PCC 7942</span></p><br />
</div><br />
<p>In this experiment, the BG-11 medium is used and the initial cell concentration is adjusted to an OD 730 of 0.2 as the last test. In addition, the sodium sulfide is diluted from 5mM to 5/16mM, while DCMU 0.25μM is added to every well.<br /><br />
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<div align="left"><br />
<p><span class="subtitle">Sulfide oxidation in Escherichia coli expressing sulfide-quinone reductase gene</span></p><br />
</div><br />
<p>Both wild type and sqr expressing Escherichia coli was cultivated in LB medium with streptomycin. The initial cell concentration was controlled to an OD600 of 0.2 and grew in the test tube inside incubator of 37 degrees Celsius and 200 rpm. Sodium sulfide was diluted from 200μM to 125μM and added into test tube. The consumption of sulfide was measured every 90 minutes by the methylene blue method (Trüper and Schlegel 1964).<br /><br />
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<h2 class="drawer-handle open">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymiq2.html
Team:NYMU-Taipei/ymiq2.html
2012-10-27T01:48:36Z
<p>Jayms: </p>
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<div class="title">Methods</div><br />
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<p><span class="subtitle">Resistance of Cyanobacteria (Synechococcus SP. PCC 7002) to Sulfide compound</span></p><br />
</div><br />
<div align="left"><br />
<p>Several Cyanobacteria have Sulfide-Quinone Reductase (sqr) and thus the ability to deprive electron from sulfide compound. According to both databases of NCBI and KEGG, the sqr in Synechococcus SP. PCC 7002 shared great similarity with that of Oscillatoria limnetica, which is reported to exhibit anoxygenic photosynthesis by consumed sulfide anion. Since we planned to express sqr from Synechococcus SP. PCC 7002 in Synechococcus SP. PCC 7942 and Escherichia coli, the experiment was designed to testify the property of the sqr. DCMU was added in the medium to inhibit photosystem II, and therefore only sodium sulfide in the medium can provide electron for carbon photoassimilation. By creating different dilution of sodium sulfide, we expected that the more sodium sulfide was present, the better the cell grew. <br /><br />
<br />
<br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/5/5e/Ymiq3.png" width="469" height="276" border="0" align="center" /><br /><br />
DCMU structure and its mechanism on photosynthesis<br /><br />
http://en.wikipedia.org/wiki/File:Diuron.png <br /><br />
<br /><br />
</div><br />
<br />
<div align="left"><br />
<p><span class="subtitle">Sodium sulfide concentration and cell growth</span></p><br />
</div><br />
<p>From the previous studies, it is suggested that <em>Synechococcus SP. PCC 7002 </em>is able to metabolize sulfide compounds. We took advantage of the results in our last experiment and adjusted the concentration of DCMU to an appropriate degree. Since sulfide would become the main reducing energy for photoassimilation under the effect of DCMU, we believe the more sulfide concentration in the wells, the better cell growth would be observed.</p><br />
<div class=out style='text-align:center'><span class="out" style="text-align:center"><img src="https://static.igem.org/mediawiki/2012/2/23/Ymiq4-1.png" width="557" height="415" border="0" align="center" /></span><br /><br />
<br /><br />
<br />
<br />
<p><span class="subtitle">The effect of sodium sulfide on Synechococcus SP. PCC 7942 growth rate</span></p><br />
</div><br />
<br />
<p>After thoroughly examined the ability of sqr in Synechococcus SP. PCC 7002, we planned to conduct a series of similar experiments on Synechococcus SP. PCC 7942. Except for the cultivation medium, other growing conditions remained the same. Instinctively, the strain expressing sqr should grow better than the wile type strain. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper.</p><br />
<div class=out style='text-align:center'><span class="out" style="text-align:center"><img src="https://static.igem.org/mediawiki/2012/5/57/Ymiq4-2.png" width="573" height="299" border="0" align="center" /></span><br /><br />
<div align="left"><br />
<p><span class="subtitle">Sulfide concentration and the growth of sqr expressing strain Synechococcus SP. PCC 7942</span></p><br />
</div><br />
<div align="left"><br />
<p>It was expected that SQR expressing strain and wild type counterpart would have different growth rate under the presence of sulfide compounds. Though sulfide is naturally toxic to Synechococcus SP. PCC 7942, the strain with sqr should be able to metabolize sulfide and therefore prosper. As the result, we analyze H2S amount to detect whether sqr gene work or not. Therefore, we perform Chemical microvolume turbidimetry method to detect H2S concentration (see Sulfur Oxide Terminator part)</div><br /><br />
<br />
<div align="left"><br />
<p><span class="subtitle">Sulfide oxidation in Escherichia coli expressing sulfide-quinone reductase gene</span></p><br />
</div><br />
<div align="left"><br />
<p>Repots have it that Escherichia coli can express functional sulfide-quinone reductase (SQR). Therefore, we slightly adjusted the previous experiment and applied to the SQR gene from Synechococcus SP. PCC 7002. With methylene blue method, we would test the efficiency of SQR sulfide oxidation. Since such method involved in measurement of optical density, it is more appropriate to perform such experiment on colorless bacteria instead of engineered cyanobacteria strain.</div><br /><br />
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<h2 class="drawer-handle open">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymis8.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
</li><br />
<br />
<br />
<li class="drawer"><br />
<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
<br />
</ul><br />
</li><br />
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<br />
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<br />
<li class="drawer last"><br />
<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
</ul> <br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymiq1.html
Team:NYMU-Taipei/ymiq1.html
2012-10-27T01:48:12Z
<p>Jayms: </p>
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<div class="title">Abstract</div><br />
<div align="left"><br />
<p><span class="subtitle">Resistance of Cyanobacteria to Sulfide compound --&gt; Sulfide-Quinone Reductase</span></p><br />
</div><br />
<div align="left"><br />
<p>Several Cyanobacteria have Sulfide-Quinone Reductase (SQR) and thus the ability to deprive electron from sulfide compound. According to both databases of NCBI and KEGG, the sqr in Synechococcus SP. PCC 7002 shared great similarity with that of Oscillatoria limnetica, which is reported to exhibit anoxygenic photosynthesis by consumed sulfide anion. Since we planned to express sqr from Synechococcus SP. PCC 7002 in Synechococcus SP. PCC 7942 and Escherichia coli, the experiment was designed to testify the property of the sqr. DCMU was added in the medium to inhibit photosystem II, and therefore only sodium sulfide in the medium can provide electron for carbon photoassimilation. By creating different dilution of sodium sulfide, we expected that the more sodium sulfide was present, the better the cell grew. <br /><br />
<br />
<br />
<div class=out style='text-align:center'><br />
<br />
<img src="https://static.igem.org/mediawiki/2012/9/95/Ymiq2.gif" alt="" width="308" height="80"><img border="0" src="https://static.igem.org/mediawiki/2012/7/7e/Ymiq1.png" align="center" /><br /><br />
The special &rdquo;CO2&rdquo; infused device for cyanobacteria incubation<br /><br />
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<h2 class="drawer-handle open">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
</ul><br />
</li><br />
<br />
<br />
<li class="drawer"><br />
<h2 class="drawer-handle">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
</ul><br />
</li><br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<br />
<li class="drawer"><br />
<h2 class="drawer-handle">Denitrifying Machine</h2><br />
<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
</ul><br />
</li><br />
<br />
<br />
<li class="drawer"><br />
<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
<br />
</ul><br />
</li><br />
<br />
<br />
<br />
<br />
<br />
<br />
<li class="drawer last"><br />
<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
</ul> <br />
</li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymis7.html
Team:NYMU-Taipei/ymis7.html
2012-10-27T01:47:39Z
<p>Jayms: </p>
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<li>We’ve already created a powerful artificial cyanobacteria with useful sulfite reductase, Cys I and Dsr, which can remove SOX from environment and protect our earth.<br /><br />
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</li><br />
<li>We’ve already established sulfite reductase, Cys I and Dsr, two biobricks. The thing is they work powerfully. Among which, CysI can remove at least 30uM H2S in 48hr!!<br /><br />
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</li><br />
<li>In the future, with sulfite reductase, we can perform our project to next step- utilize the resource from Venus or even create Venusians!! </li><br />
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<p><span class="subtitle">References</span></p><br />
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<div align="left">1. HEIKE LAUE, MICHAEL FRIEDRICH et al, Dissimilatory Sulfite Reductase (Desulfoviridin) of the Taurine-Degrading, Non-Sulfate-Reducing Bacterium<br /><br />
<em>Bilophila wadsworthia </em>RZATAU Contains a Fused DsrB-DsrD Subunit, Journal of Bacteriology 183 (2001), 5, pp. 1727-1733<br /><br />
2. Tamar Barkay and Jeffra Schaefer et al, Metal and radionuclide bioremediation: issues, considerations and potentials, Ecology and industrial microbiology 2001, 4:318–323<br /><br />
3. Werner Badziong and Rudolf K. Thauer, Growth Yields and Growth Rates of. <em>Desulfovibrio vulgaris </em>(Marburg) Growing on Hydrogen plus Sulfate and Hydrogen plus Thiosulfate as the Sole Energy Sources, Arch. Microbiol. 117, 209-214 [1978]<br /><br />
4. GERRIT VOORDOUW, The Genus <em>Desulfovibrio</em>: The Centennial, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Aug. 1995, p. 2813–2819<br /><br />
5. I. Lavilla, F. Pena-Pereira et al, Microvolume turbidimetry for rapid and sensitive determination of the acid labile sulfide fraction in waters after headspace single-drop microextraction with <em>in situ </em>generation of volatile hydrogen sulfide, Analytica Chimica Acta 647 (2009) 112–116 <br /><br />
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<h2 class="drawer-handle open">Sulfur Oxide Terminator</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
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<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
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<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymis6.html
Team:NYMU-Taipei/ymis6.html
2012-10-27T01:47:19Z
<p>Jayms: </p>
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<p>1. Desulfovibrio desulfuricans is a kind of anaerobe bacteria, so it&rsquo;s important to modify suitable environment for them.<br />
<div class=out style='text-align:center'><br /><br />
<p align="center"><img src="https://static.igem.org/mediawiki/igem.org/3/31/Ymis21.gif" alt="" width="254" height="324" /><br /><br />
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In wrong way to culture Desulfovibrio desulfuricans, the mediun turn red </p><br />
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<p>&nbsp;</p><br />
<p>2. Bioinformation tools are convenient, by using them, we can save lots of time, even though they are not totally right. </p><br />
<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/igem.org/1/18/Ymis22.gif" alt="" width="402" height="188" /><br /><br />
<p align="center">NCBI blast is a kind of powerful bioinformation tool (<a href="http://blast.ncbi.nlm.nih.gov/">http://blast.ncbi.nlm.nih.gov/</a> ) </p><br />
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<p><br /><br />
3. Standard plasmid-pSB1C3 is a unified biobrick. However, if target gene with endogenous ECoRI, PstI, XbaI and SpeI site, we have to modify pSB1C3 and create several new cloning site.<br /><br />
<br /><br />
4. According to our experiment data compared to the standard curve, we logical predict our Cys I sulfite reductase can remove at least 30uM H2S in 48hr, which is quite big amount!! That is, our artificial cyanobacteria can perform bioremediation. </p><br />
<p>5. Chemical microvolume turbidimetry method is a cheap, easy H2S detection way. But, we still need to analyze H2S with GS/MS or HPLC to increase specificity.<br /><br />
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6. Due to H2S will dissolve into water, when analyzing H2S with DPDA solution, we had better detect both water layer and gas layer. <br /><br />
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<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
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<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
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</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
<ul><br />
<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<ul><br />
<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
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References</a></li><br />
</ul><br />
</li><br />
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<li class="drawer"><br />
<h2 class="drawer-handle">Cd+2 Collector</h2><br />
<ul><br />
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<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
<ul><br />
<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms
http://2012.igem.org/Team:NYMU-Taipei/ymis5.html
Team:NYMU-Taipei/ymis5.html
2012-10-27T01:46:59Z
<p>Jayms: </p>
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href="https://2012.igem.org/Team:NYMU-Taipei/ymit1">Team</a> · <br />
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<div class="title">Practical Application in Industrial Waste Detection</div><br />
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<p>To further practical use, we believe our artificial creatures have high ability to digest H2S from industrial waste and keep our environment clean. Also, it&rsquo;s a good way to evaluate whether these sulfite reductase gene have high potential to expressed on cyanobacteria or not. We cooperate with Chung Hwa Pulp Corporation and get some industrial waste water and gas. By Co-culture with our Cys I and Dsr expressing E.coli for 24hr and 48 hr, we predict that the concentration of H2S will decrease dramatically. Both gas phase and water phase of each group are analysed by Microvolume turbidimetry method (mentioned above). Finally, the results are as the same as we imagined. In other word, the H2S concentration decrease in both gas phase and water phase. We verify that our gene, Cys I and Dsr work again!! <div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/igem.org/a/ac/Ymis16.gif" alt="" width="553" height="316" /><br /><br />
<p align="center"><img src="https://static.igem.org/mediawiki/igem.org/8/8f/Ymis17.gif" width="553" height="300" /></p><br />
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Using our artificial cyanobacteria and E.coli can clean the industrial waste produced by Chung Hwa Pulp Corporation </div><br />
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<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/igem.org/d/d6/Ymis18.gif" alt="" width="554" height="417" /></div><br />
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By simply co-culturing the waste gas and water with our modified cyanobacteria and E.coli, we can clean the pollution </div></p><br />
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<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/4/41/Ymis19.png" alt="" width="557" height="331" /><br />
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In waste gas removal experiment, our pTrc-kan-Cys I and pTrc-kan-Dsr transformed E.coli have powerful ability to convert sulfite gas waste into H2S in both water and gas layer, compared to blank control pTrc-kan transformed E.coli<br /><br />
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<div class=out style='text-align:center'><img src="https://static.igem.org/mediawiki/2012/9/97/Ymis20.png" alt="" width="557" height="337" /><br /><br />
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In waste water removal experiment, our pTrc-kan-Cys I and pTrc-kan-Dsr transformed E.coli also can convert sulfite water waste into H2S in both water and gas layer, compared to blank control pTrc-kan<br /><br />
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<h2 class="drawer-handle open">Sulfur Oxide Terminator</h2><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymis1.html">Overview</a></li><br />
<li><a title="Paper-Based Research" href="https://2012.igem.org/Team:NYMU-Taipei/ymis2.html">Paper-Based Research</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymis3.html">Experiment Design</a></li> <br />
<li><a title="Result" href="https://2012.igem.org/Team:NYMU-Taipei/ymis4.html">Result</a></li> <br />
<li><a title="Practical Application in Industrial Waste Detection" href="https://2012.igem.org/Team:NYMU-Taipei/ymis5.html">Practical Applicatioin in <br /><br />
Industrial Waste Detection</a></li><br />
<li><a title="Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymis6.html">Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymis7.html">Conclusion &amp; References</a></li><br />
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<h2 class="drawer-handle">Sulfide as Energy Generator</h2><br />
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<li><a title="Abstract" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq1.html">Abstract</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq2.html">Methods</a></li><br />
<li><a title="Experiments" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq3.html">Experiments</a></li><br />
<li><a title="Results & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq4.html">Results &amp; References</a></li><li><a title="Further Experiments after Asia Jamboree" href="https://2012.igem.org/Team:NYMU-Taipei/ymiq5.html">Further Experiments after Asia Jamboree</a></li><br />
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<h2 class="drawer-handle">Denitrifying Machine</h2><br />
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<li><a title="Background" href="https://2012.igem.org/Team:NYMU-Taipei/ymin1.html">Background</a></li><br />
<li><a title="Methods" href="https://2012.igem.org/Team:NYMU-Taipei/ymin2.html">Methods</a></li><br />
<li><a title="Results" href="https://2012.igem.org/Team:NYMU-Taipei/ymin3.html">Results</a></li><br />
<li><a title="Practical Application & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymin4.html">Practical Application &amp;<br /><br />
References</a></li><br />
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<h2 class="drawer-handle">Cd+2 Collector</h2><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymic1.html">Overview</a></li><br />
<li><a title="Experiment Design" href="https://2012.igem.org/Team:NYMU-Taipei/ymic2.html">Experiment Design</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymic3.html">Methods & Materials</a></li><br />
<li><a title="Results & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymic4.html">Results & Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymic5.html">Conclusion & References</a></li><br />
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<h2 class="drawer-handle">Becoming Venusian</h2><br />
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<li><a title="Overview" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusian.html">Overview</a></li><br />
<li><a title="Introduction" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianintro.html">Introduction</a></li><br />
<li><a title="Methods & Materials" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianmm.html">Methods &amp; Materials</a></li><br />
<li><a title="Result & Discussion" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusianrd.html">Result &amp; Discussion</a></li><br />
<li><a title="Conclusion & References" href="https://2012.igem.org/Team:NYMU-Taipei/ymivenusiancr.html">Conclusion &amp; References</a></li><br />
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Jayms