O_(139)霍乱弧菌内毒素LPS基因在大肠杆菌中的克隆表达

通过玻片凝集法从Ｏ１３９霍乱弧菌基因文库中筛选出可以表达Ｏ１３９霍乱弧菌脂多糖和Ｏ抗原的重组克隆大肠杆菌ＪＭ１０９（ｐＭＧ３１０），ＤＨ５α（ｐＭＧ３２０）和ＤＨ５α（ｐＭＧ３２１）。经免疫学方法鉴定三个重组克隆表达的脂多糖或Ｏ－抗原具有与Ｏ１３９霍乱弧菌脂多糖和Ｏ抗原相同的反应原性和免疫原性，并且三个重组克隆间的反应原性及免疫原性几乎无差别。限制性内切酶ＥｃｏＲⅠ酶切重组粒ｐＭＧ３２０和ｐＭＧ３２１发现，两个Ｏ抗原基因大小相差近１ｋｂ，这说明这两个Ｏ抗原基因属于Ｏ１３９霍乱弧菌基因组ＤＮＡ的不同区域

Cloning and Expression of LPS Genes ofV.Cholerae O139 in E.Coli

ecombinant clones expressing lipopolysaccharide(LPS) and O antigen of V.cholerae O139 were detected by constructing cosmid genomic bank and plasmid genomic bank of V.cholerae O139 to develop a vaccine against V.cholerae O139 and study structure and function of O antigen genes in the future.Recombinant clones,E.coli JM109(pMG310),DH5α(pMG320) and DH5α(pMG321) expressing LPS and O antigen of V.cholerae O139 were obtained by slide agglutination test.LPS and O antigen expressed by recombinant clones had the same immunogenicity and reactogenicity as V.cholerae O139.Agarose electrophoresis pattern showed that the size of two O antigen genes were different when recombinant plasmids,pMG320 and pMG321,were digested with restriction endonuclease EcoR I, and they belonged to different DNA fragments of genome of V.cholerae O139.