猪带绦虫TSO45W-4BX与猪CD58分子在大肠埃希菌中的联合表达  

Combined Expression of TSO45W-4BX from Taenia solium and Porcine CD58 in Escherichia coli

作　　者：骆学农[1] 郑亚东[1] 窦永喜[1] 侯俊琳[1] 景志忠[1] 才学鹏[1] 

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室,兰州730046

出　　处：《中国寄生虫学与寄生虫病杂志》2006年第4期285-289,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家重大基础研究发展规划("973")项目(G1999011906)~~

摘　　要：目的以猪CD58为分子佐剂,将CD58基因与猪带绦虫疫苗候选抗原基因TSO45W-4BX联合表达,寻找新型抗猪囊尾蚴疫苗。方法分别以重组质粒pGEM-4B和pGEM-CD58为模板,PCR扩增猪带绦虫TSO45W-4BX基因和猪CD58基因,将TSO45W-4BX与酶切处理的pGEX-4T-1定向连接,转化大肠埃希菌JM109,重组质粒经鉴定正确后,在其下游酶切插入CD58基因,PCR扩增和测序证明阅读框正确后,用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE))和蛋白质印迹法(Westernblotting)分析表达产物的免疫活性。结果pGEX-4BX和pGEX-4BX/CD58分别表达Mr41000和Mr69000的融合蛋白,pGEX-4BX表达产物主要以可溶性形式存在,而pGEX-4BX/CD58却以包涵体形式存在,但两者都能被囊尾蚴病患者血清识别。结论TSO45W-4BX与CD58基因联合表达,TSO45W-4BX仍具有免疫活性。Objective To express the TSO45W-4BX of Taenia solium in combination with CD58 as a molecular adjuvant for improving the protective efficacy of the TSO45W-4BX recombinant vaccine. Methods TSO45W-4BX and porcine CD58 genes were amplified by PCR, using recombinant plasmids pGEM-4B and pGEM-CD58 as template respectively. The CD58 fragment was inserted into the recombinant plasmid pGEX-4T-1 with directly ligated TSO45W- 4BX, The transformant was induced with IPTG and followed by identifying the integrity of the recombinant containing TSO45W-4BX and porcine CD58 with PCR and sequencing. The products were analyzed by SDS-PAGE and Western blotting. Results The expression products of Mr69000 GST-4BX/CD58 and Mr41 000 GST-4BX were present mainly in the form of inclusion bodies and soluble substance respectively, and both were recognized by sera of cysticercosis patients. Conclusions The TSO45W-4BX co-expressed with porcine CD58 conserves its immune reactivity.

关 键 词：猪带绦虫 TS045W-4BX CD58 联合表达 

分 类 号：R532.331[医药卫生—内科学] R392.11[医药卫生—临床医学]