| 伪狂犬病病毒gB和gE蛋白重组表达及抗体间接ELISA检测方法的建立 |
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| 引用本文： | 张洪亮,郝占武,解倩倩,王凤雪,张志,韩先杰,单虎,温永俊.伪狂犬病病毒gB和gE蛋白重组表达及抗体间接ELISA检测方法的建立[J].中国动物传染病学报,2022(1):98-105. |
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| 作者姓名： | 张洪亮 郝占武 解倩倩 王凤雪 张志 韩先杰 单虎 温永俊 |
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| 作者单位： | 青岛农业大学动物医学院山东省新兽药创制协同创新中心青岛市兽医生物技术工程研究中心;内蒙古农业大学兽医学院农业农村部动物疾病临床诊疗技术重点实验室;中国动物卫生与流行病学中心 |
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| 基金项目： | 国家重点研发计划项目(2016YFD0500707-7);山东省重点研发计划(2019GNC106074);山东省农业重大应用技术创新项目“生猪生态循环养殖综合技术研究与应用示范”;内蒙古农业大学高层次人才引进项目(NDGCC2016-22、NDYB2018-2)。 |
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| 摘 要： | 本研究利用PCR扩增伪狂犬病病毒(PRV)Bartha-K61株gB基因核心抗原区和闵A株gE基因核心抗原区,构建了重组质粒pET-32a-gB和pET-32a-gE,并转化至大肠杆菌BL21(DE3)感受态细胞中,经IPTG诱导进行表达.SDS-PAGE分析表明,pET32a-gB蛋白分子量约为44 kDa,pET3...
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| 关 键 词： | 猪伪狂犬病病毒 gB基因 gE基因 蛋白表达 间接ELISA |
Expression of Recombinant gB and gE Proteins of Pseudorabies Virus and Development of Indirect ELISA Assay for Antibody Detection |
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| ZHANG Hongliang,HAO Zhanwu,XIE Qianqian,WANG Fengxue,ZHANG Zhi,HAN Xianjie,SHAN Hu,WEN Yongjun.Expression of Recombinant gB and gE Proteins of Pseudorabies Virus and Development of Indirect ELISA Assay for Antibody Detection[J].Chinese Journal of Animal Infectious Diseases,2022(1):98-105. |
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| Authors: | ZHANG Hongliang HAO Zhanwu XIE Qianqian WANG Fengxue ZHANG Zhi HAN Xianjie SHAN Hu WEN Yongjun |
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| Institution: | (Shandong Collaborative Innovation Center for Development of Veterinary Pharmaceuticals,Qingdao Research Center for Veterinary Biological Engineering and Technology,College of Animal Medicine,Qingdao Agricultural University,Qingdao 266109,China;Ministry of Agriculture and Rural Affairs Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Diseases,College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;China Animal Health and Epidemiology Center,Qingdao 266032,China) |
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| Abstract: | In this study,gB and gE genes of Porcine pseudorabies virus(PRV)Bartha-K61 strain were amplifi ed by PCR and cloned into pET32a vector.The recombinant plasmids pET-32a-gB and pET-32a-gE were then transformed into E.coli BL21(DE3)for protein expression with IPTG induction.SDS-PAGE analysis showed that the pET32a-gB protein had a molecular weight of about 44 kDa,and pET32a-gE protein had a molecular weight of about 41 kDa.Subsequently,recombinant gB and gE proteins were used as coating antigens for development of indirect ELISA.In this method,both recombinant proteins specifically reacted with PRV-positive serum and the detection sensitivity reached a dilution of 1:1600.The intra-and inter-assay repeatability variation coeffi cients were<10%.The indirect ELISA method developed in this study could be used for detection of PRV antibodies and differentiation of vaccination immune response from wild-type infection in the PRV eradication program. |
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| Keywords: | Porcine pseudorabies virus gB gene gE gene protein expression indirect ELISA |
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