|
|||||
|
|
| 猪繁殖与呼吸综合征病毒兰州分离株ORF3基因的克隆及杆状病毒载体的构建 | |
| 引用本文： | 赵娜,田宏,祁燕蓉,满自萍,吴锦艳,尚佑军,何生虎,刘湘涛.猪繁殖与呼吸综合征病毒兰州分离株ORF3基因的克隆及杆状病毒载体的构建[J].西北农业学报,2009,18(3):1-5. |
| 作者姓名： | 赵娜 田宏 祁燕蓉 满自萍 吴锦艳 尚佑军 何生虎 刘湘涛 |
| 作者单位： | 1. 中国农业科学院,兰州兽医研究所,国家口蹄疫参考实验室,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃兰州,730046;宁夏大学,农学院,宁夏银川,750021 2. 中国农业科学院,兰州兽医研究所,国家口蹄疫参考实验室,家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,甘肃兰州,730046 3. 宁夏大学,农学院,宁夏银川,750021 |
| 基金项目： | 国家高技术研究发展计划(863计划)，国家科技支撑计划 |
| 摘 要： | 根据猪繁殖与呼吸综合症病毒已知序列(ch-1a,AY032626)设计包含完整ORF3序列的1对引物,采用RT-PCR方法扩增PRRSV甘肃Lanzhou株的ORF3基因,将其克隆到杆状病毒载体pFastBacHTA上,经酶切鉴定、PCR鉴定筛选出阳性重组转移载体pFastBacHTA-ORF3,并对阳性质粒进行测序及序列分析;将pFastBacHTA-ORF3转化到DH10Bac感受态细胞中,与Bacmid发生位点特异性转座作用,获得重组转座子rBacmid-ORF3.成功构建了PRRSV Lanzhou株ORF3基因的杆状病毒载体,为基因工程疫苗的研制奠定了基础. |
| 关 键 词： | 猪繁殖与呼吸综合征病毒 ORF3基因 克隆 杆状病毒表达载体 |
| 收稿时间： | 2008/10/17 0:00:00 |
| 修稿时间： | 2008/12/3 0:00:00 |
Cloning of ORF3 Gene of PRRS Virus and Construction of Its Baculovirus Epression Vector |
|
| ZHAO N,TIAN Hong,QI Yanrong,MAN Ziping,WU Jinyan,SHANG Youjun,HE Shenghu and LIU Xiangtao.Cloning of ORF3 Gene of PRRS Virus and Construction of Its Baculovirus Epression Vector[J].Acta Agriculturae Boreali-occidentalis Sinica,2009,18(3):1-5. | |
| Authors: | ZHAO N TIAN Hong QI Yanrong MAN Ziping WU Jinyan SHANG Youjun HE Shenghu and LIU Xiangtao |
| Institution: | Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou Gansu 730046, China;Agriculture College of Ningxia University, Yinchuan Ningxia 750021, China;Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou Gansu 730046, China;Agriculture College of Ningxia University, Yinchuan Ningxia 750021, China;Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou Gansu 730046, China;Agriculture College of Ningxia University, Yinchuan Ningxia 750021, China;Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou Gansu 730046, China;Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou Gansu 730046, China;Agriculture College of Ningxia University, Yinchuan Ningxia 750021, China;Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Lanzhou Gansu 730046, China |
| Abstract: | According to the published genome sequences of PRRSV(ch-1a),one pair of specific primers were designed to amplify ORF3 gene of Lanzhou strain of Gansu province by RT-PCR.The amplified frag ment was cloned into pFastBacHTA donor plasmid.The recom binant pFast BacHTA was identified by digestion with endonuclease enzyme,PCR amplification and sequencing.Then the purified plasmid was transformed into DH 10Bac-host cells,producing specific transposition with Bacmid to yield the recombinant shuttle vehicles,rBacmid-ORF3.The baculovirus epression vector was successfully constructed,which would be useful for production of gene vaccination in future. |
| Keywords: | Porcine reproductive and respiratory syndromevirus ORF3 gene Cloning Baculovirus epression vector |
| 本文献已被 维普 万方数据 等数据库收录！ | |
| 点击此处可从《西北农业学报》浏览原始摘要信息 | |
| 点击此处可从《西北农业学报》下载免费的PDF全文 | |