Regulatory

Part:BBa_K3384313:Design

Designed by: Yiqing Zhang   Group: iGEM20_NJTech_China   (2020-10-26)

pprm1 Pro


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]


Design Notes

Based on the characterization results, we found that although pprm1 possesses three PREs, it has a low induced expression level which indicates a good potential for transformation. And the background expression level of this promoter is low which means it can be used to express some proteins that require lower concentrations in the early stage. Based on the literature, we learned the general effect of PREs on promoter strength. For example, an increase in the copy number of PREs can enhance the induced expression level of the promoter. And a PRE which is aligned in the direction of the promoter may be more effective than a PRE which is in the opposite direction of the promoter. pprm1 contains 3 × PREs which are in the opposite direction of the promoter. The modified pprm1 is referred to as pprm1 Pro whose 3 × PREs are in the same direction of the promoter.Fragment1 and Fragment2 were obtained by PCR amplification using pprm1-GFP-CYC1 plasmid as the template. pprm1 Pro consists of these two fragments.The sequences of Fragment1 and Fragment2 are shown in Table1.



NJTech China--Sequence of Fragment 1 2.png



Source

Saccharomyces cerevisiae

References

[1] Heiman, M. G., and Walter, P. (2000) Prm1p, a pheromone-regulated multispanning membrane protein, facilitates plasma membrane fusion during yeast mating, The Journal of cell biology 151, 719-730.

[2] Sengupta, P., and Cochran, B. H. (1990) The Pre and Pq Box Are Functionally Distinct Yeast Pheromone Response Elements, Molecular and Cellular Biology 10, 6809-6812.