| 多头带绦虫Tm16与Tm18抗原基因的克隆及原核表达 |
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| 引用本文： | 李文卉,盖文燕,姚菊霞,曲自刚,贾万忠,罗建勋,BLAGA R,付宝权.多头带绦虫Tm16与Tm18抗原基因的克隆及原核表达[J].中国预防兽医学报,2011,33(1). |
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| 作者姓名： | 李文卉 盖文燕 姚菊霞 曲自刚 贾万忠 罗建勋 BLAGA R 付宝权 |
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| 作者单位： | 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 |
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| 基金项目： | 国家十一五科技支撑计划项目，中央级公益性科研院所基本科研业务费专项项目，中国-罗马尼亚政府间科技合作项目 |
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| 摘 要： | 为原核表达Tml6和Tm18重组蛋白,本研究以自然感染羊源脑多头坳原头节基因组DNA为模板分别扩增Tml6和Tm 18杭原基因全基因片段,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-Tm 16和pGEX-Tml8.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,经谷胱甘肤琼脂糖树脂纯化得到高纯度的可溶性的GST-Tm 16及GST-Tml8重组蛋白,western blot分析表明重组蛋白均能够被兔抗GST单克隆杭体特异性识别.
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| 关 键 词： | 多头带绦虫 原核表达 |
Cloning and prokaryotic expression of Tm16 and Tm18 antigen genes from Taenia multiceps |
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| LI Wen-hui,GAI Wen-yan,YAO Ju-xia,QU Zi-gang,JIA Wan-zhong,LUO Jian-xun,BLAGA R,FU Bao-quan.Cloning and prokaryotic expression of Tm16 and Tm18 antigen genes from Taenia multiceps[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(1). |
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| Authors: | LI Wen-hui GAI Wen-yan YAO Ju-xia QU Zi-gang JIA Wan-zhong LUO Jian-xun BLAGA R FU Bao-quan |
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| Institution: | LI Wen-hui1,GAI Wen-yan1,YAO Ju-xia1,QU Zi-gang1,JIA Wan-zhong1,LUO Jian-xun1,BLAGA R,FU Bao-quan1*(1.Key Laboratory of Veterinary Parasitology of Gansu Province,Key Laboratory of Veterinary Public Health of Ministry of Agricultural,State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China,2.University of Agricultural Sciences and Veterinary Medicine,Cluj-Napoca 400372,Romania) |
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| Abstract: | In order to express Tm16 and Tm18 antigens from Taenia multiceps,the Tm16 and Tm18 genes were amplified by PCR from genomic DNA of protoscolex of Coenurus cerebralis isolated from naturally infected sheep and sequenced.The ORF encoding Tm16 and Tm18 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 showe... |
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| Keywords: | Tm16 Tm18 |
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