Difference between revisions of "Part:BBa K4129107"
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FunsTF57 is a fusion protein consisting of the DNA-binding domain LexA, the ligand sensing domain HbaR3, transactivation domain VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR3 is a longer version linker (Ottoz et. al (2014)  compared to sBAD, which was the reference sTF (Castaño-Cerezo et. al (2020)). FunsTF05 was codon optimised to <i>A. Niger </i>.
 
FunsTF57 is a fusion protein consisting of the DNA-binding domain LexA, the ligand sensing domain HbaR3, transactivation domain VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR3 is a longer version linker (Ottoz et. al (2014)  compared to sBAD, which was the reference sTF (Castaño-Cerezo et. al (2020)). FunsTF05 was codon optimised to <i>A. Niger </i>.
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LexA is a repressor that regulates the SOS response in <i>E. coli</i> (Radman. 1975). LexA binds to a specific DNA motif, namely LexO sites (Erill. et al (2003)). HbaR is a transcription factor from <i>Rhodopseudomonas palustris</i> 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 FunsTF57 contained mutant 3 of HbaR, which has the following mutations: A45V, L69A, G71K, E77M, F85G, A86G, E87G, A88M, Y96A, L97Y, N99T and A100V.
 
LexA is a repressor that regulates the SOS response in <i>E. coli</i> (Radman. 1975). LexA binds to a specific DNA motif, namely LexO sites (Erill. et al (2003)). HbaR is a transcription factor from <i>Rhodopseudomonas palustris</i> 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 FunsTF57 contained mutant 3 of HbaR, which has the following mutations: A45V, L69A, G71K, E77M, F85G, A86G, E87G, A88M, Y96A, L97Y, N99T and A100V.

Revision as of 12:24, 11 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, the ligand sensing domain HbaR3, transactivation domain VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR3 is a longer version linker (Ottoz et. al (2014) compared to sBAD, which was the reference sTF (Castaño-Cerezo et. al (2020)). FunsTF05 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 FunsTF57 contained mutant 3 of HbaR, which has 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 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)).


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]