| 猪流行性腹泻病毒变异株M和N双基因的融合表达及免疫原性分析 |
| |
| 引用本文： | 麦伟虹,蓝海恩,廖玲玲,黄焕昌,陈钊.猪流行性腹泻病毒变异株M和N双基因的融合表达及免疫原性分析[J].畜牧与兽医,2020(1):102-106. |
| |
| 作者姓名： | 麦伟虹 蓝海恩 廖玲玲 黄焕昌 陈钊 |
| |
| 作者单位： | ;1.广西畜牧研究所 |
| |
| 基金项目： | 广西科学研究与技术开发计划项目(桂科合14125008-2-26);国家科技支撑计划子课题(2014BAD13B0102)。 |
| |
| 摘 要： | 为了获得具有良好免疫原性的猪流行性腹泻病毒(PEDV)变异毒株M和N双基因融合蛋白,本研究采用RT-PCR方法分别扩增PEDV变异毒株M、N基因,利用限制性内切酶依次将其定向克隆至pMD18-T载体,将串联的N和M融合基因片段亚克隆至pGEX-KG原核表达载体,构建pGEX-NM双基因重组质粒,转化BL21表达菌,经IPTG诱导获得了以包涵体表达的N-M双基因融合蛋白,融合蛋白经Western blot检测能够被兔抗PEDV阳性血清识别,将融合蛋白免疫BALB/c小鼠制备多克隆抗体,经间接ELISA检测其抗体效价高达1∶12 800以上。本研究获得了具有良好免疫原性的N-M双基因融合蛋白,为M和N蛋白抗原表位鉴定以及结构与功能研究、PEDV变异机制与免疫机制研究、PED新型疫苗和诊断试剂研究奠定了基础。
|
| 关 键 词： | 猪流行性腹泻病毒 变异株 M基因 N基因 融合表达 免疫原性分析 |
Fusion expression and immunogenicity analysis of M and N genes of the mutant strain of porcine epidemic diarrhea virus |
| |
| MAI Weihong,LAN Hai'en,LIAO Lingling,HUANG Huanchang,CHEN Zhao.Fusion expression and immunogenicity analysis of M and N genes of the mutant strain of porcine epidemic diarrhea virus[J].Animal Husbandry & Veterinary Medicine,2020(1):102-106. |
| |
| Authors: | MAI Weihong LAN Hai'en LIAO Lingling HUANG Huanchang CHEN Zhao |
| |
| Institution: | (Guangxi Animal Husbandry Research Institute,Nanning 530001,China) |
| |
| Abstract: | In order to obtain the fusion protein of the M and N genes of the mutant PEDV mutant strain with good immunogenicity, the M and N of the PEDV were amplified by RT-PCR, and were cloned into the pMD18-T vector sequentially by the restriction endonuclease.The tandem N and M fusion gene fragments were subcloned into the pGEX-KG prokaryotic expression vector to construct pGEX-NM double gene recombinant plasmids and to transform the BL21 expression bacteria.The fusion proteins of N and M were induced by IPTG and were expressed in the inclusion bodies.The fusion proteins could be recognized by the anti-PEDV positive serum of rabbit in Western blot detection.Polyclonal antibodies were prepared by immunizing BALB/c mice with the fusion proteins of which the titer was as high as 1:12 800 detected by indirect ELISA.In this study, the N-M double gene fusion proteins with good immunogenicity were obtained, laying a foundation for identification of M and N antigen epitopes, for research on the structure and function, and the mutation and immune mechanism of PEDV, and for development of new PED vaccines and diagnostic reagents. |
| |
| Keywords: | porcine epidemic diarrhea virus mutant strain M N fusion expression immunogenicity analysis |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
