高致病性PRRSV HuN4株Nsp4的原核表达及其抗体水平检测  

Prokaryotic expression of the Nsp4 gene of highly pathogenic PRRSV and its antibody level detection

作　　者：董建国[1] 刘永刚[1] 石文达[1] 武嘉男[1] 王刚[1,2] 刘鹤[1,3] 徐明明[1,3] 荣福龙[1,3] 李丽琴[1] 田志军[1] 蔡雪辉[1] 

机构地区：[1]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/猪传染病研究室,黑龙江哈尔滨150001 [2]山东农业大学,山东泰安271018 [3]东北农业大学,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2011年第3期219-222,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：十一五国家支撑计划(2006BAD06A07)

摘　　要：为进一步研究高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)Nsp4的免疫学特性,本研究根据PRRSV HuN4株序列,设计并合成用于扩增PRRSV HuN4株Nsp4基因的引物。将扩增产物与原核表达载体pGEX-6P-1连接,转化到大肠杆菌BL21(DE3)中。Western blot试验显示,Nsp4在大肠杆菌中获得较高水平的表达,并且具有良好的反应活性。利用该蛋白建立ELISA方法,检测PRRSV HuN4株感染仔猪的不同时间内的血清,并比较抗Nsp4,囊膜蛋白和M蛋白,N蛋白的抗体水平,结果表明Nsp4抗体升高的时间晚于囊膜蛋白和M蛋白、N蛋白出现的时间,同时抗体水平也低于其他蛋白。To detection of Nsp4 antibody in porcine reproductive and respiratory syndrome virus（PRRSV） infected pigs.A pair of primers for Nsp4 gene of PRRSV was designed according to the published sequences.The Nsp4 gene was amplified,cloned into the pGEX-6P-1 and expressed in E.coli BL21（DE3）.Western blot analysis indicated that the recombinant Nsp4 could be recognized by PRRSV positive serum.The ELISA was established with the recombinant Nsp4 as coating antigen and the antibody level of Nsp4 was detected in PRRSV infected weaning piglets at different time points.Comparing with antibodies of Sac membrane protein and M protein,N protein detected with commercial ELISA kit,the results showed that the antibody of Nsp4 was delayed and the level was lower than other antibodies.

关 键 词：PRRSV NSP4 原核表达 抗原性 

分 类 号：S852.65[农业科学—基础兽医学]