猪带绦虫TSO45W-4B基因在长双歧杆菌中的表达  

Expression of Taenia solium TSO45W-4B gene in bifidobacterium longum

作　　者：周必英[1] 刘美辰[1] 杨凤娇[1] 

机构地区：[1]贵州省遵义医学院寄生虫学教研室,遵义563003

出　　处：《重庆医科大学学报》2015年第2期243-246,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:81160206);贵州省教育厅资助项目(编号:2010040)

摘　　要：目的:在成功构建猪带绦虫大肠杆菌-双歧杆菌穿梭表达质粒p GEX-TSO45W-4B的基础上,研究猪带绦虫TSO45W-4B基因在长双歧杆菌中的表达情况。方法:将猪带绦虫大肠杆菌-双歧杆菌穿梭表达质粒p GEX-TSO45W-4B电转化入长双歧杆菌,异丙基-β-D-硫代半乳糖苷(isopropyl-β-d-thiogalactoside,IPTG)诱导表达,十二烷基硫酸钠聚丙烯酰胺凝胶电泳(sodium dodecyl sulfatepolyacrylamide gel electrophoresis,SDS-PAGE)和Western blot分析表达情况。结果:酶切、聚合酶链反应(polymerase chain reaction,PCR)和测序证实,重组质粒p GEX-TSO45W-4B成功转入长双歧杆菌。SDS-PAGE显示,重组蛋白相对分子质量(Mr)约为40 k D,经谷胱甘肽转移酶(glutathione S-transferase,GST)亲和层析后可获得纯度为85%以上的重组蛋白。Western blot显示,重组蛋白能被兔抗TSO45W-4B血清、囊虫病猪血清和囊虫病患者血清所识别。结论:猪带绦虫TSO45W-4B基因能够在长双歧杆菌中获得表达,表达的重组蛋白具有特异的抗原性。Objective：To observe the expression of the gene of TSO45W-4B of Taenia solium in bifidobacterium longum（B.longum）on the basis of successful construction of the escherichia coli（E.coil）-bifidobacterium shuttle plasmid p GEX-TSO45W-4B of Taenia solium. Methods：The shuttle expression plasmid p GEX-TSO45W-4B of Taenia solium was electroporated into B.longum. After induction with isopropyl-β-d-thiogalactoside,the expression of the recombinant protein was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis（SDS-PAGE） and Western blot. Results：Restriction analysis,polymerase chain reaction and sequencing showed that the recombinant plasmid p GEX-TSO45W-4B was successfully transformed into B.longum. As demonstrated by SDSPAGE,the relative molecular mass（Mr）of the expressed recombinant protein was approximately 40 k D,and the purity of the recombinant protein was 85% after purification withglutathione S-transferase affinity chromatography. The rabbit antiserum of TSO45W-4B,cysticercosis swine serum and cysticercosis patients serum could bind to the recombinant protein in Western blot assay. Conclusion：The gene of TSO45W-4B of Taenia solium could be expressed in B.longum and the expressed recombinant protein shows specific antigenicity.

关 键 词：猪带绦虫 TSO45W-4B 长双歧杆菌 表达 

分 类 号：R383.33[医药卫生—医学寄生虫学]