Difference between revisions of "Part:BBa K4044011"
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The role of this part is genetical regulating systems based on Gal4 as there are no other Gal4 dependant promoters presented in bacteria. When binding to promoter Gal4 domain inhibits transcription activity completely. | The role of this part is genetical regulating systems based on Gal4 as there are no other Gal4 dependant promoters presented in bacteria. When binding to promoter Gal4 domain inhibits transcription activity completely. | ||
| − | The promoter contains the sequence of CGG-N11-CCG between GACA-box and TATA-box, which can be recognized by Gal4 protein. Eleven nucleotides in the | + | The promoter contains the sequence of CGG-N11-CCG between GACA-box and TATA-box, which can be recognized by Gal4 protein. Eleven nucleotides in the core location are pj23119 promoter nucleotides. When binding to promoter Gal4 domain inhibits transcription activity completely. This promoter was tested to be active in E.coli and comparable with Anderson promoter pj23119 baseline transcriptional activity. Promoters were tested in construction containing YFP under promoter that is tested with strong RBS used (BBa_J34801). The results of QPAS1-Gal4 inhibition show a significant decrease of fluorescence and result in a close to zero level of transcription. |
fluorescence graf* | fluorescence graf* | ||
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We also have shown with a computer simulation that HTH domains of Gal4 binds exactly to CGG-N11-CCG site of the promoter. | We also have shown with a computer simulation that HTH domains of Gal4 binds exactly to CGG-N11-CCG site of the promoter. | ||
| − | + | [[File:T--LMSU--Modeling-Gal4-DNA-binding.png|400px|thumb|center|Gal4 HTH-domain binding to 5’-CGG-N11-CCG-3' promoter region of DNA]] | |
| + | [[File:T--LMSU--Modeling-otherGal4-DNA.gif|400px|thumb|center|Molecular dynamics of Gal4 HTH-domain binding to 5’-CGG-N11-CCG-3' promoter region of DNA]] | ||
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Revision as of 09:04, 21 October 2021
p119_G4-n11 (Pj23119-based partially modified Gal4 dependant promoter)
The role of this part is genetical regulating systems based on Gal4 as there are no other Gal4 dependant promoters presented in bacteria. When binding to promoter Gal4 domain inhibits transcription activity completely. The promoter contains the sequence of CGG-N11-CCG between GACA-box and TATA-box, which can be recognized by Gal4 protein. Eleven nucleotides in the core location are pj23119 promoter nucleotides. When binding to promoter Gal4 domain inhibits transcription activity completely. This promoter was tested to be active in E.coli and comparable with Anderson promoter pj23119 baseline transcriptional activity. Promoters were tested in construction containing YFP under promoter that is tested with strong RBS used (BBa_J34801). The results of QPAS1-Gal4 inhibition show a significant decrease of fluorescence and result in a close to zero level of transcription.
fluorescence graf*
We also have shown with a computer simulation that HTH domains of Gal4 binds exactly to CGG-N11-CCG site of the promoter.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 30
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]