Team:UNITN-Trento/Parts

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        <a href="eth">Ethylene</a>
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        <a href="mesa">Methyl salicylate</a>
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        <a href="blue">Blue light</a>
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        <a href="bacillus">B. subtilis</a>
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             <span class="title"><a id="bacillo">Bacillus</a></span>
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             <span class="title"><a id="bacillus"><i>Bacillus subtilis</i></a></span>
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Revision as of 16:07, 20 September 2013

Ethylene Methyl salicylate Blue light B. subtilis
Ethylene
      BBa_K1065000 EFE
Ethylene Forming Enzyme gene (EFE) that forms ethylene starting from 2-oxoglutarate.
      BBa_K1065002 EFE with double-terminator
EFE followed by the double terminator BBa_B0015.
      BBa_K1065001 EFE arabinose-regulated
This is EFE followed by a double terminator (B0015) under the control of the araCpBAD inducible promoter.
Methyl salicylate
      BBa_K1065100 BSMT1
This part encodes the BSMT1, it is the enzyme that methylates salycilic acid to obtain methyl salycilate. It’s the original part from the Eau de coli MIT iGEM 2006 project, resubmitted by us in pSB1C3.
      BBa_K1065103 PchBA
PchA ,the enzyme catalyzing salicylate biosynthesis isochorismate, and PchB that is the isochorismate-pyruvate lyase for that form the precursor for the salycilic acid.
      BBa_K1065104 SAM synthetase
This part encodes the SAM (S-AdoMet) synthetase gene, (which we friendly called "uncle SAM") . It is a methyl donor required to produce MeSA and other methylated molecules.
      BBa_K1065101 BSMT1 arabinose-regulated
A methyltransferase (BSMT1) regulated by the araCpBAD inducible promoter.
      Bba_K1065105 SAM synthetase IPTG-regulated
This is EFE followed by a double terminator (B0015) under the control of the araCpBAD inducible promoter.
      BBa_K1065102 PchBA IPTG-regulated and BSMT1 arabinose-regulated
This is EFE followed by a double terminator (B0015) under the control of the araCpBAD inducible promoter.
      BBa_K1065106 BSMT1, PchBA and SAM synthetase
An improved version of the wintergreen device with BMST1, PCHBA (BBa_K1065102) with added SAM synthetase enzymes to boost wintergreen production.
Blue light
      Bba_K1065301 amilCP IPTG-regulated
We attached pLac promoter to amilCP gene. This part does not work (sadly we saw no blue color). In fact, we started from BBa_K592009 which lacks the RBS (it is a coding part). No RBS, no party!
      Bba_K1065300 BBa_K952003 with Yf1 and FixJ IPTG-regulated.
pLac+Yf1-FixJ-pFixK2-amil GFP without RBS and with pLac upstream.
      BBa_K1065303 Yf1+FixJ+pFixK2
The starting point to build a blue-light sensing circuit: the photosensor, the response regulator with its promoter. This part was build merging BBa_K592016 and BBa_K592006.
      BBa_K1065310 Yf1+FixJ+pFixK2+Inverter+pLambda+amilCP
This is a complete blue-light sensing circuit, inverted with cI. Yf1 and FixJ are under the Anderson promoter (BBa_J23100). pLambda regulates the expression of the reporter gene amilCP. When exposed to blue-light, amilCP is synthesized, when the blue-light is not present, amilCP is not produced.
      BBa_K1065302 BBa_K1065300 improved
The same as BBa_K1065300: we performed a mutagenesis to add the missing RBS, now this works!!! It is: pLac+Yf1+FixJ+pFixK2+amilGFP.
      BBa_K1065305 BBa_K952003 improved
The same as BBa_K952003: we performed a mutagenesis to add the missing RBS, now this works!!! It is: Yf1+FixJ+pFixK2+amilGFP.
      BBa_K1065306 J23100+Yf1+FixJ
Yf1 and FixJ under the Anderson promoter (BBa_J23100).
      BBa_K1065204 pXyl+EFE
EFE in pXyl vector, specific for B. subtilis.
      BBa_K1065309 BBa_K1065302+BBa_K1065002
Complete blue-light sensing circuit without inverter, producing both amilGFP and EFE.
Bacillus subtilis
      BBa_K1065201 pXyl+GFP
An improved version of the wintergreen device with BMST1, PCHBA (BBa_K1065102) with added SAM synthetase enzymes to boost wintergreen production.
      BBa_K1065106 BSMT1, PchBA and SAM synthetase
A B. subtilis xylose inducible plasmid (from LMU Munich iGEM 2012) to produce GFPmut3b, regulated by a strong ribosome binding site (Bba_e0840).
      BBa_K1065204 pXyl+EFE
EFE in pXyl vector, specific for B. subtilis.
      BBa_K1065311 Complete blue-light sensing device with inverter, amilCP and EFE
Our complete circuit composed by:J23100, Yf1 and FixJ, pFixK2, CI, pLambda, amilCP and EFE.
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