猪繁殖与呼吸综合征病毒Nsp7的原核表达及其多克隆抗体的制备  

Prokaryotic expression of nonstructural protein 7(Nsp7) of PRRSV and preparation of mouse antiserum against Nsp7

作　　者：刘鹤[1,2] 刘永刚[2] 石文达[2] 王淑杰[2] 武嘉男[2] 董建国[2] 荣福龙[1,2] 李丽琴[2] 徐明明[1,2] 孙刚 蔡雪辉[2] 

机构地区：[1]东北农业大学动物医学院,黑龙江哈尔滨150030 [2]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/动物疫病诊断中心,黑龙江哈尔滨150001 [3]黑龙江省动物卫生监督所,黑龙江哈尔滨150090

出　　处：《中国预防兽医学报》2011年第10期812-815,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：中国农业科学院基本科研业务费(ZZKJ201114)

摘　　要：为了解猪繁殖与呼吸综合征病毒(PRRSV)Nsp7的免疫原性,本研究将PRRSV Nsp7进行了原核表达。SDS-PAGE和western blot试验表明,Nsp7在大肠杆菌中获得大量表达,并且其具有较好的反应活性。采用Nsp7免疫小鼠制备多克隆抗体,并进行间接ELISA检测,表明多克隆抗体效价达到1∶32 000以上;间接免疫荧光试验表明,多克隆抗体能够特异性识别PRRSV的Nsp7,显示出Nsp7具有较好的免疫原性。本研究获得了高纯度可溶性的PRRSV Nsp7,并进一步制备多克隆抗体,为研究Nsp7蛋白的免疫学特性提供实验依据。To investigate the immunogenicity of porcine reproductive and respiratory syndrome virus （PRRSV） Nsp7, the Nsp7 gene was cloned and expressed in E. coli, and identified by SDS-PAGE and western bolt. Furthermore, the antiserum against Nsp7 was prepared in BALB/c mice with purified recombinant Nsp7. The titer of the murine antiserum was about 1：32,000 evaluated by ELISA. The IFA and western blot also demonstrated that the antiserum specifically reacted with the native PRRSV NspT. The result showed that the recombinant Nsp7 was able to induce high level of antiserum and it could be used for Nsp7 function study and diagnosis mothed establishment.

关 键 词：猪繁殖与呼吸综合征病毒 Nsp7 原核表达 多克隆抗体 

分 类 号：S852.65[农业科学—基础兽医学]