Part:BBa_M36805
Acetaldehyde Sensor
This device senses intracellular acetaldehyde, a co-inducer of AlcR transcription factor. Activated AlcR binds its corresponding operon of the downstream promoter, activating transcription of the actuator (not included in this part).
Our plasmid was unable to be constructed by commercial companies, due to toxicity to E. coli host cells. Most of our returned mutants had mutations in the constitutive promoter, presumably weakening the expression of AlcR. We recommend future experimenters to use a weak constitutive promoter to drive AlcR expression. Nevertheless, using a Gemini actuator, we were able to generate acetaldehyde dose-dependent GFP responses in a predictable manner from one mutant. This mutant contained a 40 G deletion in the constitutive promoter. Acetaldehyde could be used up to 0.64% v/v without causing significant cell damage. In our model, normalized fluorescence increased linearly with time, but our process may have been oxygen-limited.
Thus, this is an effective acetaldehyde sensor for various applications - but due respect should be paid to the toxicity of AlcR protein.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 872
Illegal XbaI site found at 942 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 2232 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1754
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 872
Illegal XbaI site found at 942 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 872
Illegal XbaI site found at 942
Illegal AgeI site found at 1623 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1407
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