Team:UNAM Genomics Mexico/Results

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<td id="leftcolumn2" 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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<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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<td id="leftcolumn2" 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>
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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>
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AmyE 5' +CzrA/ArsR (97)+ RBS P4 + RBS CI + RFP + TERMINATOR + Sp/Smr + TERMINATOR + AmyE 3' <br /><br />
AmyE 5' +CzrA/ArsR (97)+ RBS P4 + RBS CI + RFP + TERMINATOR + Sp/Smr + TERMINATOR + AmyE 3' <br /><br />
also,  
also,  
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AmyE 5' +CzrA/ArsR (98)+ RBS P4 + RBS CI + RFP + TERMINATOR<br /><br />
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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 />
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 />
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<td  id="contentcolumn2" align= "center"><br />[[File:UnamgenomicsresultsOr3.png |x100px]] <p>'''Sp/Smr + TERMINATOR + AmyE 3''''</p></td>
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<td  id="contentcolumn" align= "center"><br />[[File:UnamgenomicsresultsOr3.png |x100px]] <p>'''Sp/Smr + TERMINATOR + AmyE 3''''</p></td>
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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">
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<td  id="contentcolumn2" align= "center"><br />[[File:UnAMgenomicsResultados andsugartbueno.png| 870px]] <p>'''Sp/Smr + TERMINATOR + AmyE 3''''</p></td>
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<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>
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<td  id="contentcolumn2" align= "center"><br />[[File:UnamgenomcismexicoArac omega.png| 870px]] <p>'''Sp/Smr + TERMINATOR + AmyE 3''''</p></td>
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<td  id="leftcolumn3" align= "center"><br />[[File:UnamgenomcismexicoArac omega.png| 870px]] <p>'''RBS AraC + omega cassette + AmyE 3''''</p></td>
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<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
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<td  id="contentcolumn2" align= "center"><br />[[File:UnamgenomicsSweetand 2.png| 870px]] <p>'''Sp/Smr + TERMINATOR + AmyE 3''''</p></td>
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<td  id="leftcolumn3" align= "center"><br />[[File:UnamgenomicsSweetand 2.png| 870px]] <p>'''Pveg + RBS XylR'''</p></td>
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<td  id="contentcolumn2" align= "center"><br />[[File:UnamgenomicsmexicoOrtenemos.png | 870px]] <p>'''Sp/Smr + TERMINATOR + AmyE 3''''</p></td>
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<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>
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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.   
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.   
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::::::::::::::::::::::::::::::::::::::[[File:UnamgenomcisUp.png|right | 120px |link=Team:UNAM_Genomics_Mexico/Results#GENERAL_RESULTS]]
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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