Difference between revisions of "Part:BBa K4129110"
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| − | + | === FunsTF61 === | |
| + | FunsTF61 is a synthetic transcription factor (sTF). FunsTF61 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor. | ||
| + | FunsTF61 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR7, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR7 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 FunsTF61. 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 FunsTF61 carried mutant 7 of HbaR, which had the following mutations: | ||
| + | A45V, L69A, G71K, 76V, E77M, Y96A, L97F, N99T, A100V and V145M. | ||
| + | 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)). | ||
Revision as of 17:17, 9 October 2022
FunsTF61
FunsTF61 is a synthetic transcription factor (sTF). FunsTF61 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor. FunsTF61 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR7, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR7 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 FunsTF61. 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 FunsTF61 carried mutant 7 of HbaR, which had the following mutations: A45V, L69A, G71K, 76V, E77M, Y96A, L97F, N99T, A100V and V145M. 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)).