Part:BBa_K4779006
UASprm1
UASprm1, an enhancer, encompasses the region from -280bp to -101bp upstream of the prm1 start codon in Saccharomyces cerevisiae. UASprm1 comprises three adjacent consensus pheromone-responsive elements (PREs). These PREs share the same orientation but are positioned opposite to the transcription start site, with a 5-nucleotide spacing between them. Furthermore, UASprm1 incorporates multiple sequences characterized by decreased nucleosome affinity, which contributes to enhancing promoter activity. To assess the viability of UASprm1 as a pheromone-inducible element, we positioned it upstream of the robust constitutive promoter pTEF2, referred to as UASprm1-pTEF2. We evaluated the fluorescence intensity of the engineered yeast strain BY4741-pRS415-UASprm1-pTEF2-GFP-CYC1 through flow cytometry, followed by data analysis using FlowJo and GraphPad Prism software. The results demonstrated a 57.78% increase in promoter activity for UASprm1-pTEF2 when induced with 5uM pheromone. This underscores UASprm1's capacity as a pheromone-inducing element, conferring desirable pheromone responsiveness to pTEF2.
Characterization
The upstream activation element UASprm1 was concatenated in frame with tef2 promoter, named UASprm1-ptef2. Then the activity of these promoters can be quantitatively measured through flow cytometer，The results are shown in Figure. The results showed that UASprm1 could confer optimal pheromone induction ability on ptef2, and the promoter activity of UASprm1-ptef2 increased by 57.78% under the induction of 5μM pheromones.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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