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多头带绦虫六钩蚴Tm45W基因的克隆表达与免疫原性分析
引用本文:牟静,杨应东,杨光友,王颖旺,安晓雪,阳爱国,古小彬,韦雷飞,文建国,王淑贤,边尧.多头带绦虫六钩蚴Tm45W基因的克隆表达与免疫原性分析[J].中国兽医科学,2011(2).
作者姓名:牟静  杨应东  杨光友  王颖旺  安晓雪  阳爱国  古小彬  韦雷飞  文建国  王淑贤  边尧
作者单位:四川农业大学动物医学院;四川省攀枝花市农林科学研究院畜牧水产所;四川省动物疫病预防控制中心;四川省雅安市雨城区畜牧局;
基金项目:教育部“长江学者和创新团队发展计划”创新团队项目(IRT0848)
摘    要:为分析多头带绦虫Tm45W重组蛋白的免疫原性,利用RT-PCR技术首次从激活的多头带绦虫六钩蚴中扩增出多头带绦虫Tm45W基因。将扩增产物重组于原核表达载体pET32a(+)中,构建pET32a-Tm45W表达载体,转化至大肠杆菌BL21(DE3)中,经IPTG诱导表达融合蛋白;表达产物进行SDS-PAGE和Western-blot分析后,将纯化的Tm45W重组蛋白免疫小鼠,用ELISA检测其血清抗体。结果显示,Tm45W基因全长为813bp,其开放阅读框为765bp,编码255个氨基酸,与多头带绦虫新疆株多头蚴45m基因(序列号:EU326106)的部分序列同源性为90.60%;表达的重组蛋白大小为45ku,与脑多头蚴病羊血清的免疫印迹分析呈阳性反应;小鼠免疫后第1周血清中即可检测到抗Tm45W蛋白的特异性抗体,并于免疫后第35天达到高峰。结果表明Tm45W重组蛋白具有较好的反应活性,能引起机体较强的体液免疫反应。

关 键 词:多头带绦虫  六钩蚴  Tm45W基因  克隆  原核表达  免疫原性  

Cloning,prokaryotic expression and immunogenicity analysis of Tm45W gene from Taenia multiceps oncospheres
MU Jing,YANG Ying-dong,YANG Guang-you,WANG Ying-wang,AN Xiao-xue,YANG Ai-guo,GU Xiao-bin,WEI Lei-fei,WEN Jian-guo,WANG Shu-xian,BIAN Yao.Cloning,prokaryotic expression and immunogenicity analysis of Tm45W gene from Taenia multiceps oncospheres[J].Veterinary Science in China,2011(2).
Authors:MU Jing  YANG Ying-dong  YANG Guang-you  WANG Ying-wang  AN Xiao-xue  YANG Ai-guo  GU Xiao-bin  WEI Lei-fei  WEN Jian-guo  WANG Shu-xian  BIAN Yao
Institution:MU Jing1,YANG Ying-dong2,YANG Guang-you1,WANG Ying-wang1,AN Xiao-xue1,YANG Ai-guo3,GU Xiao-bin1,WEI Lei-fei2,WEN Jian-guo2,WANG Shu-xian1,BIAN Yao4(1.College of Veterinary Medicine,Sichuan Agricultural University,Ya'an 625014,China,2.Animal Science and Fisheries Research Institute,Panzhihua Academy of Agricultural and Forestry Sciences,Panzhihua 617061,3.Preventive and Control Center for Animal Disease of Sichuan Province,Chengdu 610041,4.Animal Husbandry Bureau of Yucheng District of Ya'an P...
Abstract:In order to analyze the immunogenicity of Tm45W protein of Taenia multiceps,the Tm45W cDNA fragment was amplified by RT-PCR from the hatched and activated oncospheres of T.multiceps.Then,the Tm45W gene was cloned into pET32a(+) vector.The recombinant vector pET32a-Tm45W was transformed into Escherichia coli BL21(DE3) and expressed with IPTG induction.The recombinant protein was analyzed by SDS-PAGE and Western-blot.Mice were immunized with the purified recombinant protein,and anti-Tm45W antibodies were meas...
Keywords:Taenia multiceps  oncosphere  Tm45W gene  cloning  prokaryotic expression  immunogenicity  
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