| 表达牛呼吸道合胞体病毒G蛋白基因的重组Ⅰ型牛疱疹病毒的构建与鉴定 |
| |
| 引用本文： | 冯军科,朱远茂,任宪刚,祖立闯,李娇,史鸿飞,高欲燃,童光志,薛飞.表达牛呼吸道合胞体病毒G蛋白基因的重组Ⅰ型牛疱疹病毒的构建与鉴定[J].中国预防兽医学报,2010,32(2). |
| |
| 作者姓名： | 冯军科 朱远茂 任宪刚 祖立闯 李娇 史鸿飞 高欲燃 童光志 薛飞 |
| |
| 作者单位： | 中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室/大动物传染病研究室,黑龙江,哈尔滨,150001 |
| |
| 基金项目： | 黑龙江省十一五科技攻关项目 |
| |
| 摘 要： | 为构建表达牛呼吸道合胞体病毒(BRSV)G蛋白基因的牛疱疹病毒Ⅰ型(BHV-1)重组病毒,本研究将人工合成的BRSV全长G蛋白基因编码序列插入到巨细胞病毒(CMV)启动子之下构建TK基因缺失转移载体。利用磷酸钙-DNA沉淀法将该转移载体与亲本病毒BHV-1/TK-/LacZ+的基因组DNA共转染牛鼻甲细胞后收获增殖的病毒。通过反向蚀斑筛选,得到重组病毒BHV-1/TK-/G+。PCR检测结果证实G蛋白基因已经插入到了亲本病毒BHV-1/TK-/LacZ+的基因组中,间接免疫荧光试验和western blot证实BHV-1/TK-/G+中的G蛋白基因在感染的细胞中获得了表达。本研究为研制BRSV及其他重要牛传染病的BHV-1病毒活载体疫苗奠定了基础。
|
| 关 键 词： | 牛呼吸道合胞体病毒 G蛋白基因 牛疱疹病毒Ⅰ型 磷酸钙-DNA沉淀法 |
Construction and identification of a recombinant BHV-1 expressing the G protein gene of bovine respiratory syncytial virus |
| |
| FENG Jun-ke,ZHU Yuan-mao,REN Xian-gang,ZU Li-chuang,LI Jiao,SHI Hong-fei,GAO Yu-ran,TONG Guang-zhi,XUE Fei.Construction and identification of a recombinant BHV-1 expressing the G protein gene of bovine respiratory syncytial virus[J].Chinese Journal of Preventive Veterinary Medicine,2010,32(2). |
| |
| Authors: | FENG Jun-ke ZHU Yuan-mao REN Xian-gang ZU Li-chuang LI Jiao SHI Hong-fei GAO Yu-ran TONG Guang-zhi XUE Fei |
| |
| Institution: | FENG Jun-ke,ZHU Yuan-mao,REN Xian-gang,ZU Li-chuang,LI Jiao,SHI Hong-fei,GAO Yu-ran,TONG Guang-zhi,XUE Fei (Division of Livestock Infectious Diseases,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences,Harbin 150001,China) |
| |
| Abstract: | To construct a recombinant bovine herpesvirus 1 (BHV-1) expressing the G protein gene of bovine respiratory syncytial virus (BRSV), a BHV-1 transfer vector with flanking BHV-1 TK gene was constructed by inserting a synthetic G protein gene sequence of BRSV under the control of CMV promoter. The transfer vector and the parental recombinant virus (BHV-1/TK-/Lac Z+) DNA were co-transfected into BT cells using calcium phosphate-mediated transfection. The recombinant BHV-1 (designated as BHV-1/TK-/G+) was obtain... |
| |
| Keywords: | bovine respiratory syncytial virus (BRSV) G protein gene bovine hepervirus-1 calcium phosphate-mediated transfection |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
