Difference between revisions of "Part:BBa K4304008"
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===Construction of the gene ipaH containing plasmids=== | ===Construction of the gene ipaH containing plasmids=== | ||
We send the company to synthesize the DNA fragments of ipaH, and inserted it into the pUC57 vector, the sequenced data showed that the plasmids which were provided by the company were successfully constructed (Figure 1). The constructed plasmids were contained in E. coli strains, we streak inoculated them on LB solid medium plates containing corresponding antibiotics, and incubate them at 37℃ overnight, then we extracted the plasmids. | We send the company to synthesize the DNA fragments of ipaH, and inserted it into the pUC57 vector, the sequenced data showed that the plasmids which were provided by the company were successfully constructed (Figure 1). The constructed plasmids were contained in E. coli strains, we streak inoculated them on LB solid medium plates containing corresponding antibiotics, and incubate them at 37℃ overnight, then we extracted the plasmids. | ||
| − | [[File:T -- YkPaO--BBa K4304008-figure1. | + | [[File:T -- YkPaO--BBa K4304008-figure1.png|500px|thumb|center|Figure 1. the sequencing data of ipaH DNA fragment.]] |
Revision as of 12:27, 26 September 2022
ipaH Plasmid
ipaH Plasmid
Contribution
Shigella is a class of non-spore-forming Gram-negative pathogens whose pathogenicity depends on the secretion system. Shigella ipaH family proteins are co-encoded by virulence large plasmids and genes on chromosomes. We designed this plasmid by inserting ipaH DNA fragment into the plasmid pUC57 with a T7 promoter and terminator in front and after the sequence. This plasmid is used to obtain the gene fragment of Shigella which will bind with sgRNAs and recruit cas12a protein to cleave the DNA fragment of it.
Construction of the gene ipaH containing plasmids
We send the company to synthesize the DNA fragments of ipaH, and inserted it into the pUC57 vector, the sequenced data showed that the plasmids which were provided by the company were successfully constructed (Figure 1). The constructed plasmids were contained in E. coli strains, we streak inoculated them on LB solid medium plates containing corresponding antibiotics, and incubate them at 37℃ overnight, then we extracted the plasmids.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 355
Illegal SapI.rc site found at 73
Illegal SapI.rc site found at 964
Illegal SapI.rc site found at 1552