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新孢子虫SRS2基因的克隆及原核表达
引用本文:邢小勇,王权,温峰琴,郝宝成,项海涛,胡永浩. 新孢子虫SRS2基因的克隆及原核表达[J]. 动物检疫, 2012, 0(9): 43-45,59
作者姓名:邢小勇  王权  温峰琴  郝宝成  项海涛  胡永浩
作者单位:[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院上海兽医研究所,上海200241 [3]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州730050
摘    要:为构建表达新孢子虫重组蛋白SRS2原核表达系统,本研究以新孢子虫基因组为模板PCR扩增SRS2蛋白部分基因,获得大小约为998bp的目的片段,并克隆到大肠杆菌PET-32a载体中,IPTG诱导重组大肠杆菌,通过SDS—PAGE和免疫印迹分析表达产物并鉴定其生物学活性。经SDS—PAGE分析,表达的重组SRS2蛋白分子量约为54kD,免疫印迹结果表明,表达的重组蛋白与牛抗新孢子虫阳性血清反应形成一条特异性蛋白条带。本实验为进一步研究诊断和治疗新孢子虫病奠定了基础。

关 键 词:新孢子虫  SRS2基因  克隆  表达

Cloning and Expression of Neospora caninum SRS2 Gene
Xing Xiaoyong,Wang Quai,Wen Fengqin,Hao Baocheng,Xiang Haitao,Hu Yonghao. Cloning and Expression of Neospora caninum SRS2 Gene[J]. , 2012, 0(9): 43-45,59
Authors:Xing Xiaoyong  Wang Quai  Wen Fengqin  Hao Baocheng  Xiang Haitao  Hu Yonghao
Affiliation:1.College of Animal Medicine, Gansu Agricultrual University, Lanzhou, 730070 2.Shanghai Veterinary Research Institute CAAC, Shanghai, 200241: 3.Lanzhou Animal Husbandry and Veterinary Drug Research Institute, Lanzhou, 730050)
Abstract:To construct recombinant plasmid expressing Neospora caninum SRS2, the SRS2 gene from a Neospora caninum strain was amplified by PCR with a pair of primers designed according to the sequence in GenBank.A 998 bp gene was amplified and cloned into the vector PET-32a.The expression of SRS2 protein was induced by IPTG .SDS-PAGE analysis indicated that the expressed recombinant protein of SRS2 was about 54 kD.The Western blot analysis confirmed the recombinant protein was recognized by positive serum anti- Neospora caninum.The study provided a foundation for further study on the diagnosis and vaccine of neosporosis.
Keywords:Neosporacaninum   SRS2   Gene   clone   expressing
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