Team:UNAM Genomics Mexico/Results

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<center><h1>'''GENERAL RESULTS'''</h1></center><br />
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<h2>GENERAL RESULTS</h2><br /><br />
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For the heavy metal AND we sent to the registry four Biological Parts: CI, and the three different ArsR and CzrA metal sensing repressors binding fused promoters (97, 98, and 99). We did not design promoter 98 (iGEM Newcastle 2009 did), however the part was not in the registry. <br /><br />
-
 
+
The constructs we have up to now are: <br /><br />
-
For the heavy metal AND we sent to the registry four Biological Parts: CI, and the three different ArsR and CzrA metal sensing repressors binding fused promoters (97, 98, and 99). We did not design promoter 98 (iGEM Newcastle 2009 did) however the part was not in the registry. <br />  
+
<center>
-
The constructs we have up to now are: <br />
+
-
AmyE 5' +CzrA/ArsR (97 and 98)+ RBS P4 + RBS CI + RFP + TERMINATOR<br />
+
-
 
+
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+
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<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
<tr>
<tr>
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<td id="leftcolumn2" align="center"><br />[[File:UnamgenomicsAndmetals1.png | x100px]]<br /><br /><p>Our Team</p></td>
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<td id="rightcolumn2" align= "center"><br />[[ File:UnamgenomicsConstruccion1.png | 870px]]<br /><br /> <p>''' AmyE 5' +CzrA/ArsR (97)+ RBS P4 + RBS CI + RFP + TERMINATOR + Sp/Smr + TERMINATOR + AmyE 3' One complete AND'''</p></td>
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</tr>
 +
 
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<tr>
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<td id="rightcolumn2" align= "center"><br />[[File:UnamgenomicsAndmetals1.png | x90px]]<br /><br /> <p>'''AmyE 5' +CzrA/ArsR (98 and 99)+ RBS P4 + RBS CI + RFP + TERMINATOR'''</p></td>
</tr>
</tr>
<tr>
<tr>
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<td  id="contentcolumn2" align= "center"><p>The complete team of unam genomics mexico its composed mainly by undergrad students, 4 advisors, and 2 instructors</p></td>
 
</tr>
</tr>
</table>
</table>
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</center>
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AmyE 5' +CzrA/ArsR (97)+ RBS P4 + RBS CI + RFP + TERMINATOR + Sp/Smr + TERMINATOR + AmyE 3' <br /><br />
 +
also,
 +
AmyE 5' +CzrA/ArsR (98-99)+ RBS P4 + RBS CI + RFP + TERMINATOR<br /><br />
 +
 +
We still need to complete it with Sp/Smr + TERMINATOR + AmyE 3' (a part which we already have), do the same thing for promoter 99, and to add : <br /><br />
 +
 +
<center>
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
<tr>
<tr>
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<td id="leftcolumn2"align="center"><td>
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<td id="contentcolumn" align= "center"><br />[[File:UnamgenomicsresultsOr3.png |x100px]] <p>'''Sp/Smr + TERMINATOR + AmyE 3''''</p></td>
 +
</tr>
 +
</table>
 +
</center>
 +
AmyE 5' +CzrA/ArsR (98)+ RBS LasR + RBS CI to RFP + TERMINATOR and Sp/Smr + TERMINATOR + AmyE 3' as well.
 +
We are also trying to ligate our fused promoters to GFP in order to characterize them. <br /><br />
 +
For the arabinose-xylose AND gate we sent to the registry the omega cassette, pBad/pXyl, pVeg, and XylR.
 +
(XylR already existed in the registry, but ours is optimized for Bacillus subtilis). We characterized the omega cassette. <br /><br />
 +
The constructs we have up to now are:<br /><br />
 +
 +
<center>
 +
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
 +
<tr>
 +
<td  id="leftcolumn3" align= "center"><br />[[File:UnAMgenomicsResultados andsugartbueno.png| 870px]] <p>'''AmyE 5' + pBad/pXyl +RBS + P4 +RBS + CI + RBS + RFP + TT''''</p></td>
</tr>
</tr>
</table>
</table>
 +
</center>
-
[[File:UnamgenomicsresultsOr3.png |x100px |center]]
+
AmyE 5' + pBad/pXyl +RBS + P4 +RBS + CI + RBS + RFP + TT<br /><br />
-
We still need to complete them with Sp/Smr + TERMINATOR + AmyE 3' (a part which we already have), do do the same thing for promoter 99, and to add :  <br />
+
-
AmyE 5' +CzrA/ArsR (97 and 98)+ RBS LasR + RBS CI to  RFP + TERMINATOR and Sp/Smr + TERMINATOR + AmyE 3' as well. We are also trying to ligate our fused promoters to GFP in order to characterize them.  <br /> <br />
+
-
For the arabinose-xylose AND gate we sent to the registry the omega cassette, pBad/pXyl, pVeg, and XylR. (XylR already existed in the registry, but ours is optimized for Bacillus subtilis). We characterized the omega cassette. <br /><br />
+
<center>
 +
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
 +
<tr>
 +
<td  id="leftcolumn3" align= "center"><br />[[File:UnamgenomcismexicoArac omega.png| 870px]] <p>'''RBS AraC + omega cassette + AmyE 3''''</p></td>
 +
</tr>
 +
</table>
 +
</center>
-
The constructs we have up to now are:<br />
 
-
AmyE 5' + pBad/pXyl <br />
 
-
[[File:UnamgenomicsSweetand1.png | x70px]]
+
RBS AraC + omega cassette + AmyE 3'<br /><br />
 +
<center>
 +
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
 +
<tr>
 +
<td  id="leftcolumn3" align= "center"><br />[[File:UnamgenomicsSweetand 2.png| 870px]] <p>'''Pveg + RBS XylR'''</p></td>
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</tr>
 +
</table>
 +
</center>
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RBS AraC + omega cassette + AmyE 3'<br />
 
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[[File:UnamgenomicsSweetand3.png |x100px]]
 
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Pveg + RBS XylR (we are working hard to obtain the final construct which joins these two parts.<br /><br />
 
-
[[File:UnamgenomicsSweetand1.png | x70px]]
+
Pveg + RBS XylR (we are working hard to obtain the final construct which joins these tree parts.<br /><br />
-
For the A3-LasB (GusA reporter) OR gate we sent to the registry A3. <br />
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For the A3-LasB (GFP reporter) OR gate we sent to the registry A3.<br /><br />
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We have: <br />
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RBS - GusA, AmyE 5’ digested with S, P, AmyE 3’ digested with E, X, A3 promoter digested with S, P, LasB promoter digested with S, P, B0014 digested with E,Xand Omega cassette – AmyE 3’. <br /><br />
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[[File:UnamgenomicsresultadosOr1.png | x100px]]
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[[File:UnamgenomicsresultsOr2.png | x100px]]
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We have:<br /><br />
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<center>
 +
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
 +
<tr>
 +
<td  id="contentcolumn" align= "center"><br />[[File:UnamgenomicsmexicoOrtenemos.png | 870px]] <p>'''AmyE 5' + LasB + RBS + GFP + TT + A3 and need to ligate it with RBS + GFP + Terminator + omega cassette terminator + AmyE 3''''</p></td>
 +
</tr>
 +
</table>
 +
</center>
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AmyE 5' + LasB + RBS GusA Terminator and need to ligate it with A3 + RBS GusA Terminator + omega cassette terminator + AmyE 3' which we also already have. <br /><br />
 
 +
AmyE 5' + LasB + RBS + GFP + TT + A3 and need to ligate it with RBS + GFP + Terminator + omega cassette terminator + AmyE 3' which we also already have. <br /><br />
-
Regarding our work with Bacillus subtilis, we designed a new standard for working with it. This way future iGEM teams that wish to work with Bacillus subtilis will not have trouble transforming it. We designed a series of videos to explain how this is done. <br />
+
 
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[[Team:UNAM_Genomics_Mexico/Results/Bacillus | Video with the protocol ]] <br /> <br />
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== '''''Bacillus subtilis''''' ==
 +
<br />
 +
Regarding our work with ''Bacillus subtilis'', we designed a new standard protocol for working with it.  
 +
This way future iGEM teams that wish to work with Bacillus subtilis will not have trouble transforming it.
 +
We designed a series of videos to explain how this is done [[Team:UNAM_Genomics_Mexico/Results/Bacillus | Video with the protocol ]]; also PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM, in which we were able to observe the NANOTUBES,  just as Ben Yehuda et. al. reported. 
 +
<br /><br />
 +
::::::::::::::::::::::::::::::::::::::[[File:UnamgenomcisUp.png|right | 120px |link=Team:UNAM_Genomics_Mexico/Results#GENERAL_RESULTS]]
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 +
<center>
 +
<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
 +
<tr>
 +
<td id="rightcolumn2" align= "center"><br />[[File:UnamgenomicsREJ35 valores.jpg|800px| link=Team:UNAM_Genomics_Mexico/Notebook/Bacillus]]<p>'''PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM Rej35_valores:(A)Measurment of the length and width of the nanotubes connecting the (PY79) cells grown on solid LB medium. The scale bar represent 1 micrometer'''</p></td>
 +
</tr>
 +
</table>
 +
</center>
 +
<br /><br />

Latest revision as of 00:20, 26 October 2012


UNAM-Genomics_Mexico


GENERAL RESULTS



For the heavy metal AND we sent to the registry four Biological Parts: CI, and the three different ArsR and CzrA metal sensing repressors binding fused promoters (97, 98, and 99). We did not design promoter 98 (iGEM Newcastle 2009 did), however the part was not in the registry.

The constructs we have up to now are:


UnamgenomicsConstruccion1.png

AmyE 5' +CzrA/ArsR (97)+ RBS P4 + RBS CI + RFP + TERMINATOR + Sp/Smr + TERMINATOR + AmyE 3' One complete AND

UnamgenomicsAndmetals1.png

AmyE 5' +CzrA/ArsR (98 and 99)+ RBS P4 + RBS CI + RFP + TERMINATOR

AmyE 5' +CzrA/ArsR (97)+ RBS P4 + RBS CI + RFP + TERMINATOR + Sp/Smr + TERMINATOR + AmyE 3'

also, AmyE 5' +CzrA/ArsR (98-99)+ RBS P4 + RBS CI + RFP + TERMINATOR

We still need to complete it with Sp/Smr + TERMINATOR + AmyE 3' (a part which we already have), do the same thing for promoter 99, and to add :


UnamgenomicsresultsOr3.png

Sp/Smr + TERMINATOR + AmyE 3'

AmyE 5' +CzrA/ArsR (98)+ RBS LasR + RBS CI to RFP + TERMINATOR and Sp/Smr + TERMINATOR + AmyE 3' as well. We are also trying to ligate our fused promoters to GFP in order to characterize them.

For the arabinose-xylose AND gate we sent to the registry the omega cassette, pBad/pXyl, pVeg, and XylR. (XylR already existed in the registry, but ours is optimized for Bacillus subtilis). We characterized the omega cassette.

The constructs we have up to now are:


UnAMgenomicsResultados andsugartbueno.png

AmyE 5' + pBad/pXyl +RBS + P4 +RBS + CI + RBS + RFP + TT'

AmyE 5' + pBad/pXyl +RBS + P4 +RBS + CI + RBS + RFP + TT


UnamgenomcismexicoArac omega.png

RBS AraC + omega cassette + AmyE 3'


RBS AraC + omega cassette + AmyE 3'


UnamgenomicsSweetand 2.png

Pveg + RBS XylR


Pveg + RBS XylR (we are working hard to obtain the final construct which joins these tree parts.

For the A3-LasB (GFP reporter) OR gate we sent to the registry A3.

We have:


UnamgenomicsmexicoOrtenemos.png

AmyE 5' + LasB + RBS + GFP + TT + A3 and need to ligate it with RBS + GFP + Terminator + omega cassette terminator + AmyE 3'


AmyE 5' + LasB + RBS + GFP + TT + A3 and need to ligate it with RBS + GFP + Terminator + omega cassette terminator + AmyE 3' which we also already have.


Bacillus subtilis


Regarding our work with Bacillus subtilis, we designed a new standard protocol for working with it. This way future iGEM teams that wish to work with Bacillus subtilis will not have trouble transforming it. We designed a series of videos to explain how this is done Video with the protocol ; also PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM, in which we were able to observe the NANOTUBES, just as Ben Yehuda et. al. reported.

UnamgenomcisUp.png

UnamgenomicsREJ35 valores.jpg

PY79 cells were grown to midexponential phase, plated on LB agar, incubated for 6 hr at 37°C, and visualized by SEM Rej35_valores:(A)Measurment of the length and width of the nanotubes connecting the (PY79) cells grown on solid LB medium. The scale bar represent 1 micrometer



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