Part:BBa_K4779004
UASprm1-prm1
UASprm1-prm1 is a newly synthesized hybrid promoter, which is composed of two tandem modules, the enhancer UASprm1 and the prm1 promoter. This synthetic promoter extends the dynamic range of the native promoter by reducing background expression levels and increasing induced expression levels. The region from -280bp to -101bp upstream of the prm1 start codon were defined as UASprm1, which is pheromone-inducible. In pheromone-responsive promoters of Saccharomyces cerevisiae, such as pprm1, there are multiple putative Ste12 binding sites, defined as pheromone-responsive elements (PREs). UASprm1 contains three adjacent consensus PREs in the same orientation opposite to the transcription start site with space apart by 5 nucleotides. The fluorescence intensity of the engineered yeast strain BY4741-pRS415-UASprm1-prm1-GFP-CYC1 was measured using flow cytometry, and the data were subsequently processed with FlowJo and GraphPad Prism software. The results indicated that, compared to pprm1, the UASprm1-prm1 exhibited a 12.3% decrease in basal activity and a 12.6% increase in induced expression levels. This suggests that the synthetic UASprm1-prm1 expands the dynamic range of the promoter by reducing basal expression levels while enhancing induced expression levels.
Characterization
The upstream activation element UASprm1(BBa_K4779006) was concatenated in frame with prm1 promoter, named UASprm1-pprm1. Then the activity of these promoters can be quantitatively measured through flow cytometer. The results are shown in Figure 5-1. Compared with its corresponding core promoter prm1, the induced expression level of UASprm1-pprm1 is increased by 12.6%, while the background expression level persists in its original state.
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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