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中文摘要：
目的：构建细粒棘球绦虫重组质粒pBI-Eg95,分析其在根癌农杆菌（Agrobacterium tumefaciens,At）LBA4404株中的表达效率。方法：从细粒棘球蚴包囊中分离原头节,超声粉碎后抽提总RNA,采用RT-PCR方法扩增Eg95编码基因,将该基因定向克隆到植物表达载体pBI121中构建pBI-Eg95重组质粒;电穿孔转化A（tLBA4404）,抽提质粒进行酶切及PCR鉴定。用IPTG分别诱导rAt0、1、3、5、7、9、11和13h,用SDS-PAGE和Western blot鉴定所表达的蛋白质。结果：471bpEg95基因克隆成功。经酶切及PCR证实重组质粒pBI-Eg95成功转入A（tLBA4404）。用Bio-Rad Quantity one分析系统分析,表达产物分子质量（Mr）约为16.5kD,且能被感染细粒棘球蚴的鼠血清特异识别,表达效率为约占LBA4404菌体总蛋白的20%。结论：成功构建了细粒棘球绦虫重组质粒pBI-Eg95,且能在A（tLBA4404）中表达,为进一步研究细粒棘球绦虫转基因植物疫苗奠定了基础。
英文摘要：
Objective：To construct the recombinant plasmid pBI-Eg95 of E.granulosus,and to study its expression efficiency in the Agrobacterium tumefaciens（At） LBA4404 strain.Methods：Total RNA was extracted from hydatid cyst protoscoleces shattered by supersound.The Eg95 coding gene was amplified by RT-PCR,and then was cloned into the plant expression vector pBI121 to construct the recombinant plasmid pBI-Eg95.The recombinant plasmid was electroporated into At（LBA4404） strain.The recombinant At（rAt） was induced by IPTG for 0,1,3,5,7,9,11or13 hour,respectively.The expression product was identified by SDS-PAGE and Western blot assay.Results：The 471bp Eg95 coding gene was successfully amplified,and the recombinant pBI-Eg95 plasmid was successfully constructed,which was confirmed by restriction endonuclease digestion and PCR.The recombinant plasmid pBI-Eg95 was expressed successfully in the rAt,and the expression product with an approximately Mr of 16.5 kD could be detected by sera from mice infected by E.granulosus,and the expression efficiency was about 20 percent of the total bacterial protein.Conclusion：The recombinant plasmid could provide the basis for further research of the transgenic plant vaccine of E.granulosus.
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