Team:Groningen/Construct

From 2012.igem.org

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Our construct idea is simple: production of pigment under the regulation of rotten-meat reactive promoter. When the Bacillus subtilis senses the volatiles from the rotten meat, the rotten meat promoter becomes active thus allowing the production of downstream genes. We put pigment genes available downstream of the promoter so that the pigment would be produced when the promoter is activated.<br>
Our construct idea is simple: production of pigment under the regulation of rotten-meat reactive promoter. When the Bacillus subtilis senses the volatiles from the rotten meat, the rotten meat promoter becomes active thus allowing the production of downstream genes. We put pigment genes available downstream of the promoter so that the pigment would be produced when the promoter is activated.<br>
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We use our <i>Bacillus subtilis</i> backbone (BBa_K818000) that has <i>sacA</i> and chloramphenicol resistance gene for chromosomal integration and transformants antibiotic screening. This backbone also has <i>E. coli</i> origin of replication, so it can be amplified inside <i>E. coli</i>.<br>
We use our <i>Bacillus subtilis</i> backbone (BBa_K818000) that has <i>sacA</i> and chloramphenicol resistance gene for chromosomal integration and transformants antibiotic screening. This backbone also has <i>E. coli</i> origin of replication, so it can be amplified inside <i>E. coli</i>.<br>
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There are several idea to improve our current construct, such as: multi-colored pigment system and fine tuning of the pigment production.
There are several idea to improve our current construct, such as: multi-colored pigment system and fine tuning of the pigment production.
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Revision as of 17:09, 23 September 2012




Construct


The standard construct is a combination of biobricks.

  1. Backbone:Available in registry: BBa_K090403 (Backbone) - Single copy shuttle vector. Other option: build our own well-characterized backbone for our chassis.
  2. BBa_K116632+BBa_K116602 (Positive feedback loop) - Uses the pRe promoter regulated by C11 and terminated by B0010. Other options: BBa_J37019;BBa_K116639; BBaK116609.
  3. BBa_K274100 (Reporter) - Modified carotenoid pathway to produce red pigment.










  4. Or, using BBa_K274002 (Reporter) - Violet pigment producer.




Our construct idea is simple: production of pigment under the regulation of rotten-meat reactive promoter. When the Bacillus subtilis senses the volatiles from the rotten meat, the rotten meat promoter becomes active thus allowing the production of downstream genes. We put pigment genes available downstream of the promoter so that the pigment would be produced when the promoter is activated.


We use our Bacillus subtilis backbone (BBa_K818000) that has sacA and chloramphenicol resistance gene for chromosomal integration and transformants antibiotic screening. This backbone also has E. coli origin of replication, so it can be amplified inside E. coli.







There are several idea to improve our current construct, such as: multi-colored pigment system and fine tuning of the pigment production.


Multi-colored Pigment System


Fine Tuning of Pigment Production



Our sponsors:

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