Part:BBa_J100550
Variation in Constitutively Functioning AHL Repressible Promoter in E. coli
Our promoter was originally used in E. coli cultures that had had a lux-R cassette containing the lux-R gene inserted. The promoter was able to be repressed in the presence of the autoinducer N-3-oxohexanoyl-L-homoserine lactone, otherwise known as AHL, and the lux-R protein produced by the cassette. AHL is a quorum sensing chemical, meaning it was produced when the bacteria are in areas of high density. When AHL binds to the lux-R protein, the promoter is repressed. Researchers have studied this repressible behavior with both GFP and RFP production in the presence of different amounts of AHL. Increasing AHL levels correlated with decreasing fluorescence intensity. Because E. coli cultures do not naturally contain the lux-R gene, and because we do not have access to AHL, constitutive action of the promoter is all that can be measured. Insertion of the promoter by GGA into the GFP expressing region of our E. coli cells will cause them to transcribe the RFP gene and produce RFP instead. Because it should work constitutively, the promoter should cause the bacteria to steadily produce RFP.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 35
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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