Part:BBa_K4129108

FunsTF58

FunsTF58 is a synthetic transcription factor (sTF). FunsTF58 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor. FunsTF58 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR4, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR4 was a longer version (Ottoz et. al (2014) compared to sBAD (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 FunsTF58. 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 FunsTF57 carried mutant 4 of HbaR, which had the following mutations: A45V, L69A, G71K, E77W, F85G, A86G, E87G, A88G, A89P, Y96A, L97Y, A98M, N99T, A100V, and V142Q. Viral Protein 16 (VP16) from herpes simplex virus type 1 is a transcription factor that uses a transactivation domain to recruit the RNA polymerase II.The NLS SV40 is a small peptide sequence of PKKKRKV that enables transport of the protein to the nucleus (Garcia-Bustos et. al (1991)).