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- W2349748333 abstract "The common antigen components extracted from the isolated C.jejuni strain were examined by SDS PAGE and western blot analysis. The results showed that the 4A7 McAb probe appeared positive reaction with about 62 000 protein band of the CA.The genomic DNA of C.jejuni was partially digested with EcoRⅠ and Hind Ⅲ restriction endonuclease.Selected DNA fragments of 1 2.0 kb were isolated from agarose gels and ligated into the EcoRⅠ and Hind Ⅲ site of pBluescriptⅡKS(+).The ligation mixture was transformed into E.coli DH5α cells by the calcium chloride procedure. A genomic library of C.jejuni was constructed in pBluescriptⅡKS(+) plasmids. pET28a(+) were used for expression of specific protein. For screening the recombinanted E.coli DE3 colonies, it was transferred to nitrocellulose filters by replica blotting, the filters were then placed on new Luria Bertani agar plates supplemented with 30 mg/L of kanamycin,80 μg x gal and IPTG 80 μg per plate and incubated at 37℃.The recombinants were lysed in situ by soaking the filters in lysis buffer for 4 h and probed with the 4A7 McAb. Six recombinant E.coli clones carrying EcoRⅠ, Hind Ⅲ inserts of genomic DNA of C.jejuni in pET28a(+), expressed proteins recognized by the 4A7 McAb." @default.
- W2349748333 created "2016-06-24" @default.
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- W2349748333 date "2000-01-01" @default.
- W2349748333 modified "2023-09-25" @default.
- W2349748333 title "Construction of Genomic Library and Gene Cloning of Campylobacter jejuni Common Antigen" @default.
- W2349748333 hasPublicationYear "2000" @default.
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