新型鸭呼肠孤病毒σC蛋白的原核表达及其多克隆抗体制备

浙江农业学报

新型鸭呼肠孤病毒σC蛋白的原核表达及其多克隆抗体制备

1安徽农业大学动物科技学院，安徽合肥 230036；2中国农业科学院上海兽医研究所，上海 200241

出版日期:2014-07-25 发布日期:2014-08-06

Prokaryotic expression and polyclonal antibody preparation of σC protein of novel duck reovirus

1 College of Animal Science and Technology， Anhui Agricultural University， Hefei 230036， China;2 Shanghai Veterinary Research Institute， CAAS， Shanghai 200241， China

Online:2014-07-25 Published:2014-08-06

摘要/Abstract

摘要： 为了解新型鸭呼肠孤病毒(NDRV TH11株)σC蛋白的抗原性，采用RTPCR方法扩增了NDRV TH11株σC蛋白的编码基因，将其克隆于原核表达载体 pET30a(+)中，转化大肠杆菌 BL21(DE3)，经IPTG诱导表达HisσC重组蛋白。SDSPAGE和Western blot分析表明该重组蛋白获得高效表达，并具有较好的反应原性。以纯化的σC蛋白免疫实验兔制备了抗σC蛋白的多克隆抗体，间接 ELISA 检测结果显示其效价达 1∶20 000 以上；间接免疫荧光试验表明，多克隆抗体能够特异性识别NDRV的σC蛋白，显示出σC蛋白具有良好的免疫原性。

关键词: 新型鸭呼肠孤病毒；&sigma, C基因；原核表达；多克隆抗体

Abstract: To investigate the immunogenicity of σC protein of a novel duck reovirus (NDRV TH11)， the σC gene was cloned into pET30a(+) and expressed in E.coli BL21(DE3). SDSPAGE and Western blot analysis showed that the expressed had good immunological activity. The polyclonal antibody was obtained from the rabbit immunized with purified recombinant protein and its titer was about 1∶20 000 by detection of indirect ELISA. Indirect immunofluorescence assay also demonstrated that the polyclonal antibody reacted specially with the native NDRV σC protein. This study will be helpful for preparation of peptide vaccines and σC function study.

Key words: novel duck reovirus, σC gene, prokaryotic expression, polyclonal antibody

毕庄莉1，2，朱英奇1，2，陈宗艳2，李传峰2，孟春春2，王桂军1，刘光清2，*. 新型鸭呼肠孤病毒σC蛋白的原核表达及其多克隆抗体制备[J]. 浙江农业学报.

BI Zhuangli1，2， ZHU Yingqi1，2， CHEN Zongyan2， LI Chuanfeng2， MENG Chunchun2， WANG Guijun1， LIU Guangqing2，*. Prokaryotic expression and polyclonal antibody preparation of σC protein of novel duck reovirus[J]. .