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| 牛病毒性腹泻病毒E0基因原核表达载体的构建及表达 | |
| 引用本文： | 王璐,郭春娟,吴星星,宫玉玲,季新成,冉多良.牛病毒性腹泻病毒E0基因原核表达载体的构建及表达[J].新疆农业大学学报,2012,35(2):125-128. |
| 作者姓名： | 王璐 郭春娟 吴星星 宫玉玲 季新成 冉多良 |
| 作者单位： | 1. 新疆农业大学动物医学学院,乌鲁木齐830052;新疆出入境检验检疫局,乌鲁木齐830063 2. 新疆农业大学动物医学学院,乌鲁木齐,830052 3. 新疆出入境检验检疫局,乌鲁木齐,830063 |
| 基金项目： | 新疆维吾尔自治区高技术研究发展项目 |
| 摘 要： | 采用RT-PCR方法,利用牛病毒性腹泻病毒(BVDV)C24V株接种MDBK细胞,提取病毒RNA,扩增出BVDV-E0基因,将所得片段与pET-28a表达载体连接,转化至BL-21大肠杆菌中,筛选阳性克隆,鉴定后证明pET-28a-E0原核表达载体构建成功。经IPTG诱导后收集菌体进行SDS-PAGE和Western-blot鉴定,鉴定结果表明,目的蛋白在大肠杆菌中得以表达。 |
| 关 键 词： | 牛病毒性腹泻病病毒 E0基因 克隆 原核表达 |
Construction of Prokaryotic Expression Vector of Bovine Viral Diarrhea Virus EO Gene and Its Expression |
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| WANG Lu , GUO Chun-juan , WU Xing-xing , GONG Yu-ling , Jl Xin-cheng , RAN Duo-liang.Construction of Prokaryotic Expression Vector of Bovine Viral Diarrhea Virus EO Gene and Its Expression[J].Journal of Xinjiang Agricultural University,2012,35(2):125-128. | |
| Authors: | WANG Lu GUO Chun-juan WU Xing-xing GONG Yu-ling Jl Xin-cheng RAN Duo-liang |
| Institution: | 1(1.College of Animal Medicine,Xinjiang Agricultural University,Urumqi 830052,China;2.Entry and Exit Inspection and Quarantine Bureau of Xinjiang,Urumqi 830063,China) |
| Abstract: | C24V strains of bovine viral diarrhea virus(BVDV) were used to be inoculated into MDBK cells to extract viral RNA,to amplify BVDV-E0 gene and the fragments obtained were connected with the pET-28a expression vector to be transformed into E.coli BL-21 to screen out the posetive clones.The results showed that identification of pET-28a-E0 prokaryotic expression vector was successfully constructed.The bacteria were collected after IPTG induction by SDS-PAGE and Western-blot identification,protein identification results show that to be expressed in E.coli. |
| Keywords: | bovine viral diarrhea virus E0 gene cloning prokaryotic expression |
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