Difference between revisions of "Part:BBa K4129105"

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The synthetic transcription factor, FunsTF16 (LexA-SL-HbaR10-B112-SV40)

FunsTF16 is a synthetic transcription factor (sTF). FunsTF16 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor.

FunsTF16 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR10, transactivation domain; B112 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR was the same as sBAD (SL) (Castaño-Cerezo et. al (2020)).

LexA is a repressor that regulates the SOS response in E. coli (Radman. 1975). LexA binds to a specific DNA motif, LexO (Erill. et al (2003)), and it is the DNA binding domain that interacts with LexO that is used in FunsTF16. HbaR is a transcriptional factor from Rhodopseudomonas palustris that initiates transcription in the presence of benzoic acid or in the presence of benzoic acid derivatives (Egland. Et al (2000) (Castaño-Cerezo et. al (2020)). We created 16 mutants of HbaR and FunsTF16 carried mutant number 10 of HbaR, which had the following mutations: A45S, L64I, A86G, A88Y, Y96A and L97G. The transactivation domain B112 is from E. coli, which were experimentally proven to initiate transcription of a synthetic promoter in S. cerevisiae (Ottoz et. al (2014)). The NLS SV40 is a small peptide sequence of PKKKRKV that enables transport of the protein to the nucleus (Garcia-Bustos et. al (1991)).

Sequence and Features