多房棘球绦虫重组Bb-EmⅡ/3-Em14-3-3疫苗的构建及鉴定  

Construction and identification of the recombinant Bb-EmⅡ/3-Em14-3-3 vaccine of Echinococcus multilocularis

作　　者：杨梅[1] 李文桂[1] 朱佑明[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》2007年第10期1026-1029,共4页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(No30500423);教育部重点项目(No205131);重庆市科委攻关计划(No03-43-8);重庆市教委科研基金(NoKJ050313);重庆市卫生局科研基金(No03-2-099和05-2-203)资助项目

摘　　要：目的构建和鉴定多房棘球绦虫(Em)重组双歧杆菌(Bb)-EmⅡ/3-Em14-3-3疫苗。方法通过PCR扩增EmⅡ/3和Em14-3-3抗原编码基因,然后采用基因拼接法(geneSOEing)剪接EmⅡ/3和Em14-3-3,得到EmⅡ/3-Em14-3-3融合基因;将该融合基因定向克隆到大肠埃希菌-双歧杆菌穿梭表达载体pGEX-1λT,构建重组质粒pGEX-EmⅡ/3-Em14-3-3;用电穿孔法将该质粒导入Bb,构建多房棘球绦虫重组Bb-EmⅡ/3-Em14-3-3疫苗。结果PCR成功扩增出2554bp的EmⅡ/3-Em14-3-3融合基因;双酶切证实EmⅡ/3-Em14-3-3融合基因成功插入pGEX-1λT中;PCR证实rBb-EmⅡ/3-Em14-3-3疫苗构建成功。结论成功构建了多房棘球绦虫重组Bb-EmⅡ/3-Em14-3-3疫苗,为该疫苗的开发和利用打下了坚实的实验基础。To construct and identify the recombinant Bb-EmⅡ/3-Em14-3-3 vaccine of Echinococcus multilocularis. EmⅡ/3 and Em14-3-3 antigen gene were amplified by PCR and the fusion gene EmⅡ/3-Em14-3-3 was obtained with gene SOE-ing. Then the fusion gene was cloned into Escherichia coli-Bifidobacteria shuttle plasmid pGEX-D,T to construct pGEX-ErnⅡ/ 3-Em14-3-3. The recombinant plasmid was electroporated into Bifidobacteria bifidum （Bb） to construct rBb-EmⅡ/3-Em14-3- 3 vaccine. A 2 554bp fusion gene of EmⅡ/3-Em14-3-3 was successfully amplified by PCR and cloned into pGEX-D,T by restriction analysis, and the rBb-EmⅡ/3-Em14-3-3 vaccine was successfully constructed by PCR. In this way, the rBb-EmⅡ/3- Em14-3-3 vaccine of Echinococcus multilocularis is successfully constructed, which lays the experimental foundation of exploitation and utilization of this vaccine.

关 键 词：多房棘球绦虫 重组Bb-EmⅡ/3-Em14-3-3疫苗 构建 鉴定 

分 类 号：R383.3[医药卫生—医学寄生虫学]