Difference between revisions of "Part:BBa K4129111"

Latest revision as of 21:16, 11 October 2022

The fungal synthetic transcription factor, FunsTF62 (LexA-LL-HbaR8-VP16-SV40)

FunsTF62 is a synthetic transcription factor (sTF). FunsTF62 should initiate the transcription through the 6xLexO minimal promoter. This sTF is designed to be the sensing part of the biosensor.

FunsTF62 is a fusion protein consisting of the DNA-binding domain from LexA, the ligand sensing domain from HbaR8, the transactivation domain from VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR8 is a longer linker (Ottoz et. al (2014) compared to the linker used in sBAD, which was the reference sTF (Castaño-Cerezo et. al (2020)). FunsTF62 was codon optimised to A. niger.

LexA is a repressor that regulates the SOS response in E. coli (Radman. 1975). LexA binds to a specific DNA motif, namely LexO sites (Erill. et al (2003)). HbaR is a transcription factor from Rhodopseudomonas palustris that initiates transcription in the presence of benzoic acid (Egland. et al (2000) or in the presence of benzoic acid derivatives (Castaño-Cerezo et. al (2020)). We created 16 mutants of HbaR and FunsTF62 carried mutant 8 of HbaR, which had the following mutations: A45V, L69A, G71K, E77A, A86G, E89G, A90G, A91P, Y96A, L97Y, A98S, N99T, A100V and V145Y.

Viral Protein 16 (VP16) from Herpes simplex virus type 1 is a transcription factor with a transactivation domain that recruits RNA polymerase II (Hirai et al. (2010)).The NLS SV40 is a small peptide sequence of PKKKRKV that enables transport of the protein to the nucleus (Garcia-Bustos et. al (1991)).

The intented function was not proven in A. niger.

Sequence and Features