猪带绦虫重组BCG-TSOL18疫苗构建及其表达  

Construction and expression of a recombinant BCG-TSOL18 vaccine of Taenia solium

作　　者：杨凤娇[1] 周必英[1] 

机构地区：[1]遵义医学院寄生虫学教研室,遵义563000

出　　处：《中国人兽共患病学报》2015年第10期952-956,962,共6页Chinese Journal of Zoonoses

基　　金：贵州省科技厅社会发展攻关项目(黔科合SY字[2011]3038号)~~

摘　　要：目的构建猪带绦虫重组BCG-TSOL18疫苗,研究TSOL18基因在BCG中的表达情况。方法通过酶切的方法从重组质粒pGEX-TSOL18获取猪带绦虫TSOL18基因,将其定向克隆到大肠杆菌-分枝杆菌穿梭表达质粒pMV261中,构建猪带绦虫重组质粒pMV261-TSOL18,进行酶切、PCR和测序鉴定;再将其电穿孔转化入BCG,构建猪带绦虫重组BCGTSOL18疫苗,进行PCR鉴定;SDS-PAGE和Western blot分析TSOL18基因在BCG中的表达情况。结果通过酶切成功获得了393bp的TSOL18基因片段;酶切、PCR和测序鉴定证明成功构建了猪带绦虫重组质粒pMV261-TSOL18;PCR鉴定证实猪带绦虫重组质粒pMV261-TSOL18成功转入BCG中,提示猪带绦虫重组BCG-TSOL18疫苗构建成功;SDS-PAGE分析发现在相对分子质量(Mr)约为14.7kD处有明显的TSOL18目的蛋白条带,Western blot证实表达的TSOL18目的蛋白能被兔抗血清和囊虫病猪血清所识别。结论成功构建了猪带绦虫重组BCG-TSOL18疫苗,TSOL18基因能够在BCG中成功表达,表达的TSOL18重组蛋白具有特异的抗原性,为该疫苗的进一步研究奠定了基础。We constructed a recombinant Bacillus Calmette-Guerin（BCG）-TSOL18 vaccine of Taenia solium and observed the expression of the TSOL18 gene in BCG. The TSOL18 gene of Taenia soliurn was obtained from the recombinant plasmid pGEX-TSOL18 by digestion method and cloned into Escherichia coli （E. cold mycobacterium shuttle plasmid pMV261 to con- struct the recombinant plasmid pMV261-TSOL18 of Taenia solium, and the recombinant plasmid was identified by restriction enzyme digestion, PCR and DNA sequencing. Then, the recombinant plasmid was transformed into BCG by electroporation to construct the recombinant BCCr-TSOL18 vaccine of Taenia solium, and the vaccine was identified by PCR. The expression of the TSOL18 gene in BCG was identified by SDS-PAGE and Western blot. The 393 bp TSOL18 gene fragment was successfully obtained by restriction enzyme digestion. Restriction enzyme digestion, PCR and DNA sequencing suggested that the recombi- nant plasmid pMV261-TSOL18 of Taenia solium was successfully constructed. PCR confirmed that the recombinant plasmid pMV261-TSOL18 of Taenia soliurn was successfully transformed into BCG, suggesting that the recombinant BCG-TSOL18 vaccine of Taenia solium was successfully constructed. SDS PAGE showed that the relative molecular mass （Mr） of TSOL18 target protein was approximately 14.7 kD. Results of western blot showed the TSOL18 target protein could be recognized by rabbit antiserum or cysticereosis swine serum. The recombinant BCG-TSOL18 vaccine of Taenia solium was successfully con- structed. The TSOL18 gene of Taenia solium was successfully expressed in BCG and the expressed TSOL18 recombinant pro- tein had specific antigenicity. This result would lay a foundation for further study of the vaccine.

关 键 词：猪带绦虫 重组BCGTSOL18疫苗 构建 表达 

分 类 号：R383.33[医药卫生—医学寄生虫学]