Difference between revisions of "Part:BBa K4129107"

Revision as of 21:23, 10 October 2022

The fungal synthetic transcription factor, FunsTF57 (LexA-LL-HbaR3-VP16-SV40)

FunsTF57 is a synthetic transcription factor (sTF). FunsTF57 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor.

FunsTF57 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR3, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR3 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 FunsTF57. 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 3 of HbaR, which had the following mutations: A45V, L69A, G71K, E77M, F85G, A86G, E87G, A88M, Y96A, L97Y, N99T and A100V.

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)).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 673
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 607
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 765
1000
COMPATIBLE WITH RFC[1000]

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 __NOTOC__
+<partinfo>BBa_K4129107 short</partinfo>
+FunsTF57 is a synthetic transcription factor (sTF).  FunsTF57 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor.
+FunsTF57 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR3, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR3 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 <i>E. coli</i> (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 FunsTF57. 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 3 of HbaR, which had the following mutations: A45V, L69A, G71K, E77M, F85G, A86G, E87G, A88M, Y96A, L97Y, N99T and A100V.
-=== FunsTF57 ===
-FunsTF57 is a synthetic transcription factor (sTF).  FunsTF57 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor.
-FunsTF57 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR3, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR3 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 FunsTF57. 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 3 of HbaR, which had the following mutations: A45V, L69A, G71K, E77M, F85G, A86G, E87G, A88M, Y96A, L97Y, N99T, A100V.
 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)).
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