Difference between revisions of "Part:BBa K3384314:Design"
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===Design Notes===
 
===Design Notes===
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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 specific 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. We doubled the 3 × PREs sequence in natural pprm1. The modified pprm1 is referred to as pprm1 Ultra which contains 6 × PREs.
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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 specific 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. We doubled the 3 × PREs sequence in natural pprm1. The modified pprm1 is referred to as pprm1 Ultra which contains 6 × PREs. Fragment1 and Fragment3 were obtained by PCR amplification using pprm1-GFP-CYC1 plasmid as the template. pprm1 Ultra consists of these two fragments. The sequences of Fragment1 and Fragment3 are shown in Table1.
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<p> </p>
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[[File:NJTech_China--Sequence_of_Fragment_1_3.png|width='100%' valign='top'| |center|thumb|550px|''<b>Table 1 </b>The sequence of Fragment1 and Fragment3.]]
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===Source===
 
===Source===

Revision as of 03:37, 27 October 2020

pprm1 Ultra


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 specific 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. We doubled the 3 × PREs sequence in natural pprm1. The modified pprm1 is referred to as pprm1 Ultra which contains 6 × PREs. Fragment1 and Fragment3 were obtained by PCR amplification using pprm1-GFP-CYC1 plasmid as the template. pprm1 Ultra consists of these two fragments. The sequences of Fragment1 and Fragment3 are shown in Table1.



Table 1 The sequence of Fragment1 and Fragment3.



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.