作　　者： ; ; ; ; ;

机构地区： 西北农林科技大学动物科技学院

出　　处： 《动物医学进展》 2011年第5期77-81,共5页

摘　　要： PCR扩增淋球菌CMCC29400、CMCC29403株孔蛋白(PI)A、B及表位基因,使用linker GS-GGSG并通过重叠PCR法将表位基因连接在PIA、PIB上形成融合基因PIA-表位、PIB-表位,分别与克隆载体pMD-18T连接,测序正确后将目的基因插入表达载体pET32a(+)中,构建pET32a-PIA-表位、pET32a-PIB-表位、pET32a-PIA和pET32a-PIB重组表达质粒,PCR、双酶切鉴定及测序正确后转化大肠埃希菌BL21(DE3),IPTG诱导蛋白表达,经SDS-PAGE鉴定分析,成功表达融合蛋白。 Porin I and lipid oligosaccharide epitopes（Loe） of Neisseria gonorrhoeae are poteneial anti-gonorrhea vaccines.Porin I contains PIA and PIB.The prokaryotic expression vector of the fusion genes PIA-Loe and PIB-Loe was constructed,and the fusion proteins PIA-Loe and PIB-Loe were expressed.The PIA and PIB genes were amplified from the standard strains of Neisseria gonorrhoeae by PCR,and the Loe genes were synthetized.Fragment PIA,PIB and fragment Loe were fused with the linker GSGGSG by overlapping PCR,and a 1 392 bp and a 1 455 bp fragments of the fusion genes PIA-Loe and PIB-Loe were found on the agarose gel.Then the fusion genes PIA-Loe and PIB-Loe were inserted into pET-32a for expression in E.coli BL21（DE）3.The SDS-PAGE analysis showed that fusion proteins PIA-Loe and PIB-Loe were successfully expressed,which laid a foundation for developing a Neisseria gonorrhoeae vaccine.

关 键 词： 淋球菌 孔蛋白 脂寡糖表位 基因融合 表达

领　　域： [生物学]