Team:UNAM Genomics Mexico/Results

From 2012.igem.org

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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 />
We have: <br />
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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RBS - GFP, 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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<table border="0"  height="150" cellspacing="15" bgcolor="transparent" id="tablecontentbg" cellpadding="10">
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<td id="leftcolumn3" align= "center"><br />[[File:UnamgenomicsresultadosOr1.png | x100px]]<p>'''AmyE 5' + LasB + RBS GusA Terminator'''</p></td>
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<td id="leftcolumn3" align= "center"><br />[[File:UnamgenomicsresultadosOr1.png | x100px]]<p>'''AmyE 5' + LasB + RBS GFP Terminator'''</p></td>
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<td id="contentcolumn" align= "center"><br />[[File:UnamgenomicsresultsOr2.png | x100px]]<p>'''A3 + RBS GusA Terminator + omega cassette terminator + AmyE 3'''</p></td>
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<td id="contentcolumn" align= "center"><br />[[File:UnamgenomicsresultsOr2.png | x100px]]<p>'''A3 + RBS GFP Terminator + omega cassette terminator + AmyE 3'''</p></td>
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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 />
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AmyE 5' + LasB + RBS GFP Terminator and need to ligate it with A3 + RBS GFP Terminator + omega cassette terminator + AmyE 3' which we also already have. <br /><br />

Revision as of 06:42, 25 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:
AmyE 5' +CzrA/ArsR (97 and 98)+ RBS P4 + RBS CI + RFP + TERMINATOR


UnamgenomicsAndmetals1.png

AmyE 5' +CzrA/ArsR (97 and 98)+ RBS P4 + RBS CI + RFP + TERMINATOR


UnamgenomicsresultsOr3.png

Sp/Smr + TERMINATOR + AmyE 3'

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 :
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.

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:
AmyE 5' + pBad/pXyl


UnamgenomicsSweetand1.png

AmyE 5' + pBad/pXyl


RBS AraC + omega cassette + AmyE 3'



UnamgenomicsSweetand3.png

RBS AraC + omega cassette + AmyE 3'

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


UnamgenomicsSweetand1.png

RBS AraC + omega cassette + AmyE 3'

For the A3-LasB (GFP reporter) OR gate we sent to the registry A3.
We have:
RBS - GFP, 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’.



UnamgenomicsresultadosOr1.png

AmyE 5' + LasB + RBS GFP Terminator


UnamgenomicsresultsOr2.png

A3 + RBS GFP Terminator + omega cassette terminator + AmyE 3


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


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.
Video with the protocol