Difference between revisions of "Part:BBa K4129110"
Magnus Haahr (Talk | contribs) |
Magnus Haahr (Talk | contribs) |
||
| (2 intermediate revisions by the same user not shown) | |||
| Line 2: | Line 2: | ||
<partinfo>BBa_K4129110 short</partinfo> | <partinfo>BBa_K4129110 short</partinfo> | ||
| − | FunsTF61 is a synthetic transcription factor (sTF). | + | FunsTF61 is a synthetic transcription factor (sTF). FunsTF61 should initiate the transcription through the 6xLexO minimal promoter. This sTF is designed to be the sensing part of a biosensor. |
| − | FunsTF61 is a fusion protein consisting of the DNA-binding domain LexA, the ligand sensing domain HbaR7, transactivation domain VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR7 is a longer | + | FunsTF61 is a fusion protein consisting of the DNA-binding domain from LexA, the ligand sensing domain from HbaR7, the transactivation domain from VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR7 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)). FunsTF61 was codon optimised to <i>A. niger </i>. |
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 FunsTF61 carried mutant 7 of HbaR, which had the following mutations: | 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 FunsTF61 carried mutant 7 of HbaR, which had the following mutations: | ||
| Line 11: | Line 11: | ||
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)). | 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. | + | The intented function was not proven in <i>A. niger</i>. |
<!-- --> | <!-- --> | ||
Latest revision as of 21:15, 11 October 2022
The fungal synthetic transcription factor, FunsTF61 (LexA-LL-HbaR7-VP16-SV40)
FunsTF61 is a synthetic transcription factor (sTF). FunsTF61 should initiate the transcription through the 6xLexO minimal promoter. This sTF is designed to be the sensing part of a biosensor.
FunsTF61 is a fusion protein consisting of the DNA-binding domain from LexA, the ligand sensing domain from HbaR7, the transactivation domain from VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR7 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)). FunsTF61 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 FunsTF61 carried mutant 7 of HbaR, which had the following mutations: A45V, L69A, G71K, E77M, Y96A, L97F, N99T, A100V and V145M.
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 673
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 607
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 765
- 1000COMPATIBLE WITH RFC[1000]