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中文摘要：
目的构建和鉴定细粒棘球绦虫（Eg）重组双歧杆菌（Bb）-Eg95-EgA31融合基因疫苗。方法自行设计引物，从细粒棘球蚴包囊中分离原头节，超声粉碎后提取总RNA为模板，通过RT—PCR分别扩增Eg95和EgA31抗原编码基因，然后采用基因拼接法（gene SOEing）剪接Eg95和EgA31，得到Eg95-EgA31融合基因，经BamH I和EcoR I双酶切，定向克隆到大肠杆菌-双歧杆菌穿梭表达载体pGEX-1λT中，转化大肠杆菌BL21（DE3）感受态细胞，构建重组质粒pGEX-Eg95-EgA31，抽提质粒进行双酶切鉴定，电穿孔法转化两歧双歧杆菌（Bifidobacteriabifidum，Bb），构建细粒棘球绦虫重组Bb-Eg95-EgA31融合基因疫苗，抽提质粒进行PCR扩增鉴定。结果RT-PCR扩增出约1016bp的Eg95-EgA31融合基因；重组质粒用双酶切鉴定可切出预期大小片段，以具有氨苄青霉素抗性的rBb中抽提的质粒为模板进行PCR扩增可得到约1016bp的Eg95-EgA31融合基因片段。结论成功构建了细粒棘球绦虫重组Bb-Eg95-EgA31融合基因疫苗，为该疫苗的开发利用奠定了实验基础。
英文摘要：
In this study, the total RNA was extracted from hydatid cyst protoscoleces shattered by ultrasound breaking and the gene encoding Eg95-EgA31 antigen was amplified by RT-PCR from template of the total RNA using the primers designed according to DNA sequence of Eg95 and EgA31. The Eg95-EgA31 fusion gene was obtained with gene SOEing , cloned to E. coli-Bifidobacteria shuttle plasmid pGEX-1λT and transformed into E. coli BL21（DE3） competent cells to construct plasmid pGEX-Eg95-EgA31 by using BamHI and EcoRI. This recombinant plasmid was identified by restriction endonuelease digestion and electroporated into Bifidobacterium bifiduzn to construct fusion gene vacciner rBb-Eg95-EgA31. The extracted plasmid was identified by PCR. Experimental result showed that the fusion gene Eg95-EgA31 of 1 016 bps in length was amplified by RT-PCR and the size of the products obtained by restriction endonuclease digestion was just the same as expected. It is evident that the recombinant gene vacciner Bb-Eg95-EgA31 of Echinococcus granulosus has been successively constructed and identified, which may lay experimental foundation for this vaccine.
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