Part:BBa_K2365515

Mutant of TEF1

Mutant of TEF1 Yeast promoter; Constitutive expression

Transcription elongation factor gene promoter of Saccharomyces cerevisiae constitutive expression promoter in the commonly used system. According to the element and registry and database of synthetic biology database bioparts for synthetic biology (http://npbiosys.scbit.org/regulatoryelementdetails/00007) design and synthesis of tef1 mutations.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 166
  

                                          Fluorescence intensity was measured by flow cytometry

Measurements of specific fluorescence were performed using cells harvested from the logarithmic phase during growth in shake flasks.

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.

Contribution

• Group:Jiangnan_China iGEM team 2021
• Author:Shiyu Yu

Saccharomyces cerevisiae is a typical eukaryote, so it is very important for us to choose a suitable promoter to express genes.

In our project, we need to express three genes on one plasmid. In the process of consulting the literature, we found an article called "A Cas9 based toolkit to program gene expression in Saccharomyces cerevisiae" [1], which described in detail the relative strength of different promoters in different media and culture time. In the follow-up experiments, we referred to this literature, selected the promoter of appropriate intensity, and achieved good results. So we share this article here, hoping to give a reference to our friends who haven't read this article yet.

Reference

[1]Reider Apel A, d'Espaux L, Wehrs M, Sachs D, Li RA, Tong GJ, Garber M, Nnadi O, Zhuang W, Hillson NJ, Keasling JD, Mukhopadhyay A. A Cas9-based toolkit to program gene expression in Saccharomyces cerevisiae. Nucleic Acids Res. 2017 Jan 9;45(1):496-508. doi: 10.1093/nar/gkw1023. Epub 2016 Nov 28. PMID: 27899650; PMCID: PMC5224472.