Difference between revisions of "Part:BBa K2365051"
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| − | TPI1 Yeast promoter;Triose-phosphateisomerase, triosephosphate isomerase promoter. Constitutive expression; come from Saccharomyces cerevisiae BY4742 genomic sequence . | + | TPI1 Yeast promoter;Triose-phosphateisomerase, triosephosphate isomerase promoter. Constitutive expression; come from Saccharomyces cerevisiae BY4742 genomic sequence .It plays an important role in glycolysis, is essential for the energy generation, which has been found in almost all organisms, including mammals, insects, fungi, plants and most bacteria. |
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[[File:TPI1 TPI1-CYC1 NAU-05.jpeg|350px|center]] | [[File:TPI1 TPI1-CYC1 NAU-05.jpeg|350px|center]] | ||
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| + | Nature promoter from Saccharomyces cerevisiae genome amplification obtained constitutive promoter, the original gene are associated with the metabolism of yeast.And PCR amplification the cyc1 terminator from the genome,next can form the promoter-cyc1t fusion fragments by overlap extension. Between the elements of the paired homologous arm are reserved for standard cleavage sites, constructed on the expression vector prs423 and inserted GFP (S65T)in the fusion fragments , the recombinant plasmid tansform into yeast sey6210 and test 440-550nm fluorescence intensity. | ||
| + | [[File:启动子全部.jpeg|400px|center]] | ||
| + | Figure.The fluorescence of the four test devices and control transformed into Saccharomyces cerevisiae and inoculated in YPD broth was measured after 8h. | ||
| + | Nevertheless, the attractiveness of yeast as a platform organism (especially for metabolic engineering applications) needs continued efforts to expand the set of tools available. | ||
Latest revision as of 00:50, 2 November 2017
TPI1 promotor
TPI1 Yeast promoter;Triose-phosphateisomerase, triosephosphate isomerase promoter. Constitutive expression; come from Saccharomyces cerevisiae BY4742 genomic sequence .It plays an important role in glycolysis, is essential for the energy generation, which has been found in almost all organisms, including mammals, insects, fungi, plants and most bacteria.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 47
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 119
Nature promoter from Saccharomyces cerevisiae genome amplification obtained constitutive promoter, the original gene are associated with the metabolism of yeast.And PCR amplification the cyc1 terminator from the genome,next can form the promoter-cyc1t fusion fragments by overlap extension. Between the elements of the paired homologous arm are reserved for standard cleavage sites, constructed on the expression vector prs423 and inserted GFP (S65T)in the fusion fragments , the recombinant plasmid tansform into yeast sey6210 and test 440-550nm fluorescence intensity.
Figure.The fluorescence of the four test devices and control transformed into Saccharomyces cerevisiae and inoculated in YPD broth was measured after 8h. Nevertheless, the attractiveness of yeast as a platform organism (especially for metabolic engineering applications) needs continued efforts to expand the set of tools available.