Regulatory

Part:BBa_K2365051

Designed by: Tong Cheng   Group: iGEM17_NAU-CHINA   (2017-10-18)


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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 47
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 119


TPI1 TPI1-CYC1 NAU-05.jpeg

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.

启动子全部.jpeg

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.

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