Part:BBa_K4129109

FunsTF59

FunsTF59 is a synthetic transcription factor (sTF). FunsTF59 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor. FunsTF59 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR5, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR5 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 FunsTF59. 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 FunsTF59 carried mutant 5 of HbaR, which had the following mutations: A45V, L69A, G71K, 76V, E77M, M79L, A86G, E87G, A88M, Y89A,L97P,N99T,A100V, V145Y and K148Y. 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)).