|
|||||
|
|
| 猪繁殖与呼吸综合征病毒ORF5基因的克隆及杆状病毒表达载体的构建 | |
| 引用本文： | 赵娜,田宏,吴锦艳,满自萍,尚佑军,何生虎,刘湘涛.猪繁殖与呼吸综合征病毒ORF5基因的克隆及杆状病毒表达载体的构建[J].华北农学报,2009,24(1). |
| 作者姓名： | 赵娜 田宏 吴锦艳 满自萍 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,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,国家口啼疫参考实验室,甘肃,兰州;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,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,国家口啼疫参考实验室,甘肃,兰州 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) |
| 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ORF5序列的1对引物,采用RT-PCR方法扩增PRRSV甘肃LZH株的ORF5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-ORF5,并对阳性质粒进行测序及序列分析;将pFastBacHTA-ORF5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-ORF5;并成功构建了PRRSV LZH株ORF5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 ORF5基因 克隆 杆状病毒表达 |
Cloning of ORF5 Gene of PRRS Virus and Construction of Its Baculovirus Expression Vector |
|
| ZHAO Na,TIAN Hong,WU Jin-yan,MAN Zi-ping,SHANG You-jun,HE Sheng-hu,LIU Xiang-tao.Cloning of ORF5 Gene of PRRS Virus and Construction of Its Baculovirus Expression Vector[J].Acta Agriculturae Boreali-Sinica,2009,24(1). | |
| Authors: | ZHAO Na TIAN Hong WU Jin-yan MAN Zi-ping SHANG You-jun HE Sheng-hu LIU Xiang-tao |
| Institution: | 1.Chinese Acadeny of Agriculture Science;State Key Laboratory of Veterinary Etiological Biology;Key Laboratory of Animal Virology of Ministy of Agriculture;National FMD Reference of Ministy of Agriculture;National FMD Reference Laboratory;Lanzhou 730046;China;2.College of Agronomy;Ningxia University;Yinchuan 750021;China |
| Abstract: | According to the published genome sequences of PRRSV,one pair of specific primers were designed to amplify ORF5 gene of LZH strain of GANSU province by RT-PCR.The amplified fragment was cloned into pFastBacHTA donor plasmid.The recombinant pFastBacHTA was identified by digestion with endonuclease enzyme,PCR amplification and sequencing.Then the purified plasmid was transformed into DH10Bac-host cells,producing specific transposition with Bacmid to yield the recombinant shuttle vehicles,rBacmid-ORF5.The bacu... |
| Keywords: | Porcine reproductive and respiratory syndrome virus ORF5 gene Cloning Baculovirus expression |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |