多头带绦虫Tm16与Tm18抗原基因的克隆及原核表达  

Cloning and prokaryotic expression of Tm16 and Tm18 antigen genes from Taenia multiceps

作　　者：李文卉[1] 盖文燕[1] 姚菊霞[1] 曲自刚[1] 贾万忠[1] 罗建勋[1] BLAGA R 付宝权[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/农业部兽医公共卫生重点开放实验室/甘肃省动物寄生虫病重点实验室,甘肃兰州730046 [2]University of Agricultural Sciences and Veterinary Medicine,Cluj-Napoca 400372,Romania

出　　处：《中国预防兽医学报》2011年第1期76-79,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家十一五科技支撑计划(2007BAD40B04);中央级公益性科研院所基本科研业务费专项(BRF080303);中国-罗马尼亚政府间科技合作项目(39-18)

摘　　要：为原核表达Tm16和Tm18重组蛋白,本研究以自然感染羊源脑多头蚴原头节基因组DNA为模板分别扩增Tm16和Tm18抗原基因全基因片段,测序鉴定后合成其开放阅读框架DNA片段,将基因组序列中的内含子去除并对其稀有密码子进行改造优化,构建重组表达质粒pGEX-Tm16和pGEX-Tm18。转化大肠杆菌BL21后以IPTG诱导表达,SDS-PAGE分析表达产物并进行纯化。结果显示:在大肠杆菌中表达出带有GST标签的大小约为39.6 ku和39.4 ku的重组蛋白,经谷胱甘肽琼脂糖树脂纯化得到高纯度的可溶性的GST-Tm16及GST-Tm18重组蛋白,western blot分析表明重组蛋白均能够被兔抗GST单克隆抗体特异性识别。In order to express Tml6 and Tml8 antigens from Taenia multiceps, the Tml6 and Tml8 genes were amplified by PCR from genomic DNA of protoscolex of Coenurus cerebralis isolated from naturally infected sheep and sequenced. The ORF encoding Tml6 and Tml8 proteins were synthesized with the optimized codes to replace the rare codens and cloned into pGEX-4T-1 prokaryotic expression vector. The recombinant plasmid was transformed into E. coli BL21 and the expression products were analyzed by SDS-PAGE. The results showed the recombinant Tml6 and Tml8 genes of T. multiceps were expressed as 39.6 ku and 39.4 ku protein, respectively. The soluble fusion proteins were purified using GST resin and could be detected by western blot using the rabbit monoclonal anti-GST antibody.

关 键 词：多头带绦虫 Tm16 Tm18 原核表达 

分 类 号：S852.7[农业科学—基础兽医学]