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Team:Cambridge/Bacillus
From 2008.igem.org
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'''Building on the work of the last year's Cambridge iGEM team, we are exploring applications of the gram-positive chassis ''B.subtillis''. Easy to handle and transform, this bacterium is much better to use than ''E.coli'' wherever protein import/export is concerned, e.g. in our signalling project. An important part of our effort is to establish standard protocols and parts to work in ''B.subtillis'', characterise control elements, and develop new vectors. You can find our [[IGEM:Cambridge/2008/Turing Pattern Formation/Experiments/Bacillus subtilis transfomation| ''Bacillus'' protocols here]].''' | '''Building on the work of the last year's Cambridge iGEM team, we are exploring applications of the gram-positive chassis ''B.subtillis''. Easy to handle and transform, this bacterium is much better to use than ''E.coli'' wherever protein import/export is concerned, e.g. in our signalling project. An important part of our effort is to establish standard protocols and parts to work in ''B.subtillis'', characterise control elements, and develop new vectors. You can find our [[IGEM:Cambridge/2008/Turing Pattern Formation/Experiments/Bacillus subtilis transfomation| ''Bacillus'' protocols here]].''' | ||
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==Improved GFP== | ==Improved GFP== | ||
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We created a codon-optimized and biobricked version of the 'Superfolder' GFP engineered by Pédelacq et al (2006). This GFP variant folds 4x faster and fluoresces 4x brighter than even P7 'Superfast' GFP. [[Team:Cambridge/Improved_GFP|Read more...]] | We created a codon-optimized and biobricked version of the 'Superfolder' GFP engineered by Pédelacq et al (2006). This GFP variant folds 4x faster and fluoresces 4x brighter than even P7 'Superfast' GFP. [[Team:Cambridge/Improved_GFP|Read more...]] | ||
| - | =Creating new Biobrick-compatible ''B.subtilis'' vectors= | + | ==Creating new Biobrick-compatible ''B.subtilis'' vectors== |
[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Cambridge Vectors submitted:] | [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Cambridge Vectors submitted:] | ||
We are using the In-Fusion™ PCR method from ClonTech to create new Biobrick-compatible integrative and episomal vectors. These vectors will allow us to insert Biobricks into the ''Bacillus subtilis'' genome at the AmyE locus and as a 3-5 copy plasmid that will replicate autonomously in ''Bacillus''. | We are using the In-Fusion™ PCR method from ClonTech to create new Biobrick-compatible integrative and episomal vectors. These vectors will allow us to insert Biobricks into the ''Bacillus subtilis'' genome at the AmyE locus and as a 3-5 copy plasmid that will replicate autonomously in ''Bacillus''. | ||
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| - | =='''B. subtilis Promoters Designed''' | + | =='''B. subtilis Promoters Designed'''== |
[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Cambridge Biobricks submitted:] | [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Cambridge Biobricks submitted:] | ||
* Pupp - a strong constitutive ''Bacillus cereus'' promoter present on ''B. subtilis'' vector ECE166 | * Pupp - a strong constitutive ''Bacillus cereus'' promoter present on ''B. subtilis'' vector ECE166 | ||
Revision as of 04:08, 30 October 2008
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Building on the work of the last year's Cambridge iGEM team, we are exploring applications of the gram-positive chassis B.subtillis. Easy to handle and transform, this bacterium is much better to use than E.coli wherever protein import/export is concerned, e.g. in our signalling project. An important part of our effort is to establish standard protocols and parts to work in B.subtillis, characterise control elements, and develop new vectors. You can find our Bacillus protocols here.
Improved GFP We created a codon-optimized and biobricked version of the 'Superfolder' GFP engineered by Pédelacq et al (2006). This GFP variant folds 4x faster and fluoresces 4x brighter than even P7 'Superfast' GFP. Read more... Creating new Biobrick-compatible B.subtilis vectors[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Cambridge Vectors submitted:] We are using the In-Fusion™ PCR method from ClonTech to create new Biobrick-compatible integrative and episomal vectors. These vectors will allow us to insert Biobricks into the Bacillus subtilis genome at the AmyE locus and as a 3-5 copy plasmid that will replicate autonomously in Bacillus.
Successful transformation of BacillusTransformation with episomal vectorsECE166 : High-level constitutive expression of a Green Fluorescent Protein (with the promoter Pupp) Read more...
 Transformation with integration vectorsECE153 and ECE112 : Transformation in IA751 : Amylase test  
B. subtilis Promoters Designed[http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2008&group=Cambridge Biobricks submitted:]
Read more about the constructs made... Read more about the primers used... Information about the B. subtilis vectors can be found here.
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