Difference between revisions of "Part:BBa K2559003"
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<partinfo>BBa_K2559003 short</partinfo> | <partinfo>BBa_K2559003 short</partinfo> | ||
| − | The | + | The BBa_K2559003 , PrplJ promoter is a strong endogenous promoter in Escherichia coli K 12. We used it to express single guide RNA in our CRISPR/cas9 system. |
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===Usage and Biology=== | ===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 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 | + | 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 modeling PrplJ, Pdapa and PcaiF.We 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. |
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[[File:Scau-china-2018-11.png|800px|thumb|center|Figure1 The result of the fluorescent intensity measurement ]] | [[File:Scau-china-2018-11.png|800px|thumb|center|Figure1 The result of the fluorescent intensity measurement ]] | ||
Revision as of 18:08, 15 October 2018
Ecoli promoter of sgRNA(PrplJ)
The BBa_K2559003 , PrplJ promoter is a strong endogenous promoter in Escherichia coli K 12. We used it to express single guide RNA in our CRISPR/cas9 system.
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 modeling PrplJ, Pdapa and PcaiF.We 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.
Basing on the model predition and experiment testing results,we used PrplJ as the promoter of sgRNA
The part is facilitate the in-depth research for other teams!
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]