细粒棘球绦虫重组BCG-Eg95疫苗构建及鉴定  

Construction and identification of the Echinococcus granulosus recombinant BCG-Eg95 vaccine

作　　者：朱佑明[1] 李文桂[1] 

机构地区：[1]重庆医科大学附属第一医院传染病寄生虫病研究所,重庆400016

出　　处：《中国人兽共患病学报》2006年第9期805-808,共4页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No.30200239);重庆市高等学校优秀中青年骨干教师资助项目

摘　　要：目的构建细粒棘球绦虫重组BCG-Eg95疫苗并鉴定。方法从包囊中分离原头节,超声粉碎后抽提总RNA为模板,采用RT-PCR方法扩增Eg95编码基因并鉴定。将该基因片段定向克隆到大肠杆菌-分枝杆菌穿梭表达载体pBCG中构建pBCG-Eg95重组质粒。用电穿孔法将重组质粒导入BCG菌构建rBCG-Eg95疫苗,用HgCl2筛选经PCR鉴定。结果经电泳及测序证实从原头节扩增出471bp Eg95蛋白编码基因。重组质粒pBCG-Eg95经酶切及PCR证实,并经PCR鉴定已成功转入BCG。结论成功构建细粒棘球绦虫重组BCG-Eg95疫苗,为rBCG-Eg95疫苗进一步研究奠定基础。To construct and identify the Echinococcus granulosus recombinant BCG-Eg95 vaccine. Total RNA was extracted from hydatid cyst protoscoleces shattered by suppersound, and used as template. The Eg95 coding gene was amplified by RT-PCR and identified by sequenceing and electrophoresis, and then cloned into E. coli-Mycobacteria shuttle plasmid pBCG to construct the recombinant plasmiel pBCG-Eg95, which was confirmed by restriction endonuclease digestion and PCR. This recombinant plasmid was introduced into BCG by electroporation. The rBCG positive clones were screened by HgCl2 and identified by PCR method. Thus in this way the 471bp Eg95 coding gene was successfully amplified, purely cloned into pBCG and successfully introduced into BCG. In conclusion, the Echinococcus granulosus recombinant BCG-Eg95 was successfully constructed. The results of the study could provide for the basis to further research of rBCG-Eg95 vaccine.

关 键 词：细粒棘球绦虫 重组BCG-EG95疫苗 

分 类 号：R383.33[医药卫生—医学寄生虫学]