Difference between revisions of "Part:BBa K2559003"

Revision as of 18:01, 15 October 2018

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 rplj/dapa/caif promoter.

Due to model predition and experiment test result,we use 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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 ===Usage and Biology===
-We have obtained this promoter from a database, PromEC ( http://margalit.huji.ac.il/promec/index.html) .PromEC is an updated compilation of E. coli mRNA promoter sequences. It includes documentation on the location of experimentally identified mRNA transcriptional start sites on the E. coli chromosome, as well as the actual sequences in the promoter region. The database is currently updated as of July 2000 and includes 471 entries.
+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 strength  of these promoters and tested the prplJ, pdapA, and pcaiF expression strength by comparing the fluorescent intensity.You can use PromEC  and our model to obtain more useful promoter information.
+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.
-The eGFP expression in E.coli(DH5 alpha) and was driven by prplj/pdapa/pcaif promoter.
 [[File:Scau-china-2018-11.png|800px|thumb|center|Figure1 The result of the fluorescent intensity measurement ]]
 [[File:Scau-china-2018-12.png|800px|thumb|center|Figure2 Fluorescent intensity of eGFP driven by by rplj/dapa/caif promoter.]]