Part:BBa_I766200

pSte2

endogenous promoter of Ste2

Estonia TUIT Contribution

Figure 1. Model for the STE2 gene regulation. From Yeast pheromone receptor genes STE2 and STE3 are differently regulated at the transcription and polyadenylation level, by Di Segni et al., 2011 .

Figure 2. Characterization of the endogenous pSTE2 promoter used. GFP was expressed from pSTE2 in far1∆ yeast cells. Flow cytometry was used to measure GFP fluorescence in the cells after 2 hours of incubation in each concentration of the exogenous α-factor. Black circles represent mean single-cell fluorescence. The black curve represents a sigmoidal transfer function obtained by fitting to the data. Error bars, SEM n=3. From Supplementary Materials of Secreting and Sensing the Same Molecule Allows Cells to Achieve Versatile Social Behaviors, by (Youk & Lim, 2014) . .

STE2 promoter (pSTE2) regulates the expression of the STE2 gene coding for the G-protein-coupled receptor for α-factor pheromone. The model for the STE2 gene regulation was proposed by Di Segni et al. in 2011 (Figure 1). According to the model, the STE2 gene is activated in yeast cells by Mcm1. In this case, the canonical poly(A) site in the 3′ UTR is used (long and filled arrow). On the other side, several proteins are involved in the repression of the STE2 promoter in α cells (Mcm1, α2, Ssn6, Tup1, and others).

The expression of the gene under the STE2 promoter was analyzed in the work of Youk & Lim where they studied yeast cell-to-cell communication (Youk & Lim, 2014). Researchers expressed GFP reporter under pSTE2 promoter and analyzed the correlation of the GFP fluorescence in the far1∆ yeast cells with different α-factor concentrations (Figure 2).

Methodology

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
  COMPATIBLE WITH RFC[1000]