Difference between revisions of "Part:BBa K1937003:Design"
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The BBa_K1937003 part was designed to demonstrate the induction of pNagA by NAG. It belongs tothe antifungal module in the Paleolitis project of iGEM Toulouse 2016 (http://2016.igem.org/Team:Toulouse_France).
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<b>=Part: BBa_K1937003 (pNagA)=</b>
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The part include the pNagA promoter and its RBS (position 3,595,156 to 3,595,356 of the <i>Bacillus</i> genome). The promoter controls the expression of the RFP fluorescent reporter gene. A NheI restriction site has been placed before the RFP ORF to facilitate promoter swapping.
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(Chassis E. coli, carrier plasmid pSB1C3, part destined for use in <i>Bacillus subtilis</i>)
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Length: 1035 bp<br>
  
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<b>Background:</b>
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pNagA is the promoter of the NagA gene, which expresses a N-acetylglucosamine-6-phosphate deacetylasein in <i>Bacillus subtilis</i> (http://www.biocyc.org/gene?orgid=BSUB&id=BSU35010).
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In <i>Bacillus subtilis</i>, this promoter has been reported to induce expression in presence of N-acetylglucosamine 6-phosphate (Bertram et al., 2011: https://www.ncbi.nlm.nih.gov/pubmed/21602348). This NAG molecule is the major component of chitin, a characteristic molecule of fungi.
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This BioBrick is a part developed by the Toulouse 2016 iGEM team (http://2016.igem.org/Team:Toulouse_France)
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<b>This part:</b>
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The BBa_K1937003 part was designed to demonstrate the induction of pNagA by NAG. It belongs to the antifungal module in the Paleolitis project of iGEM Toulouse 2016 (http://2016.igem.org/Team:Toulouse_France).
 +
The part include the pNagA promoter and its RBS (position 3,595,156 to 3,595,356 of the Bacillus genome). The promoter controls the expression of the RFP fluorescent reporter gene. A NheI restriction site has been placed before the RFP ORF to facilitate promoter swapping.
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<br>
 
[[File:BBa_K1937003_-construction.png]]
 
[[File:BBa_K1937003_-construction.png]]
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<b>Validation:</b>
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The part was sub-cloned in the pSBBsOK-mini plasmid (BBa_K1937001) for validation (resulting in part BBa_K1937004). In presence of N-acetyl glucosamine in the media, the clones are expected to be red, and white without this substrate Nag.

Revision as of 12:29, 17 October 2016

=Part: BBa_K1937003 (pNagA)= (Chassis E. coli, carrier plasmid pSB1C3, part destined for use in Bacillus subtilis) Length: 1035 bp

Background: pNagA is the promoter of the NagA gene, which expresses a N-acetylglucosamine-6-phosphate deacetylasein in Bacillus subtilis (http://www.biocyc.org/gene?orgid=BSUB&id=BSU35010). In Bacillus subtilis, this promoter has been reported to induce expression in presence of N-acetylglucosamine 6-phosphate (Bertram et al., 2011: https://www.ncbi.nlm.nih.gov/pubmed/21602348). This NAG molecule is the major component of chitin, a characteristic molecule of fungi. This BioBrick is a part developed by the Toulouse 2016 iGEM team (http://2016.igem.org/Team:Toulouse_France)

This part: The BBa_K1937003 part was designed to demonstrate the induction of pNagA by NAG. It belongs to the antifungal module in the Paleolitis project of iGEM Toulouse 2016 (http://2016.igem.org/Team:Toulouse_France). The part include the pNagA promoter and its RBS (position 3,595,156 to 3,595,356 of the Bacillus genome). The promoter controls the expression of the RFP fluorescent reporter gene. A NheI restriction site has been placed before the RFP ORF to facilitate promoter swapping.
BBa K1937003 -construction.png

Validation: The part was sub-cloned in the pSBBsOK-mini plasmid (BBa_K1937001) for validation (resulting in part BBa_K1937004). In presence of N-acetyl glucosamine in the media, the clones are expected to be red, and white without this substrate Nag.