Regulatory
P-rpij

Part:BBa_K2559003

Designed by: Yinpin Huang   Group: iGEM18_SCAU-China   (2018-10-07)
Revision as of 18:03, 15 October 2018 by Weiteng (Talk | contribs) (Usage and Biology)


Ecoli promoter of sgRNA(PrplJ)

The rpij promoter is a strong endogenous promoter in Escherichia coli K 12;we use it to start the expression of single guide RNA


Usage and Biology

We obtained this promoter from a database named PromEC ( http://margalit.huji.ac.il/promec/index.html) .PromEC is an updated compilation of E. coli mRNA promoter sequences. It includes detailed information of genesites which have been experimentally identified mRNA transcriptional starting position in the E. coli chromosome, as well as the actual sequences of the promoters. We have built up a model to measure the expression intensity of these promoters. Eventually, we selected three promoters including: PrplJ, Pdapa and PcaiF based on the results of modelingPrplJ, Pdapa and PcaiFWe further made chimeric fluorescent fusion proteins between the promoter and GFP respectively and tested the fluorescent intensity driven by different promoters. The combination of PromEC and our modeling can be used as a efficient tool to figure out the information about interested promoters. The GFP expression in E.coli (DH5 α) and was driven by PrplJ, Pdapa and PcaiF promoter.

Figure1 The result of the fluorescent intensity measurement
Figure2 Fluorescent intensity of eGFP driven by by PrplJ, PdapA, PcaiF promoter.

Due to model predition and experiment test result,we used PrplJ as the promoter of sgRNA

The part is facilitate the in-depth research for other teams!


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]


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