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- W2365892010 abstract "The fed operon gene cluster encoding the F18ab-serotype fimbriae was amplified by Expand Long Template PCR using the genomic DNA template from Shiga-like toxigenic E.coli reference strain 107/86.The predicted PCR products with the restriction enzyme sites at each end were digested and then cloned into the expression vector pBR322,the recombinant plasmid pBR322-fed with the inserts of the whole fed gene clusters were constructed and screened,further confirmed by the means of combination with both restriction endonuclease analysis and sequencing.The fimbriae F18ab were isolated and purified from the recombinant E.coli SE5000(pBR322-fed),and only a single major band of protein with size of approximately 15 000 was visualized in Coomassie blue-stained gels after SDS-PAGE,and this protein band was recognized positively by the polyclonal sera against fimbriae F18ab in Western-blotting assay.The results in agglutination assay showed that the polyclonal sera,purified IgG,and monoclonal antibody(Mab) directed against the major subunit of F18ab fimbriae(FedF),F18ab fimbriae from the reference strain E.coli 107/86 reacted to the recombinant E.coli SE5000 expressing the fimbriae F18ab,respectively.Small intestine epithelial cells from the susceptible piglets with the genotypes of FUT1 gene M307GG were prepared and tested for the adherence to the recombinant E.coli SE5000(pBR322-fed) under the microscopic examination.Adhesion and adhesion inhibition test showed the recombinant E.coli SE5000(pBR322-fed) could adhere to the jejunal epithelial cells in vitro as the reference strain E.coli 107/86 did.The polyclonal anti-sera directed against fimbriae F18ab can efficiently inhibit the fimbriae-mediated susceptible piglet jejunal epithelial cells adherence to the recombinant E.coli SE5000(pBR322-fed) and reference strain E.coli 107/86." @default.
- W2365892010 created "2016-06-24" @default.
- W2365892010 creator A5046259508 @default.
- W2365892010 date "2012-01-01" @default.
- W2365892010 modified "2023-09-25" @default.
- W2365892010 title "Cloning and expression of F18 fimbrial operon gene cluster from Shiga-like toxigenic E.coli(SLTEC) using non-induced expression system" @default.
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