犬瘟热病毒N基因原核表达及多克隆抗体的制备  

PROKARYOTIC EXPRESSION OF N GENE OF CANINE DISTEMPER VIRUS AND PREPARATION OF POLYCLONAL ANTIBODIES

作　　者：李丹丹[1,2] 刘柱[1,2] 丁明洋 李传峰[2] 陈宗艳[2] 张淼涛[1] 刘光清[2] 

机构地区：[1]西北农林科技大学动物医学院,杨凌712100 [2]中国农业科学院上海兽医研究所,上海200241 [3]甘肃农业大学动物医学院,兰州530005

出　　处：《中国动物传染病学报》2016年第3期1-5,共5页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金项目(31270194;31502068);上海市科技兴农重点攻关项目(2016043);上海市科委创新项目(13391901602);公益性农业科研专项(201303046);中央级公益性科研院所基本科研业务费专项(2016JB01;31101848)

摘　　要：采用PCR方法扩增犬瘟热病毒N基因,将其克隆至原核表达载体p ET-32a(+)中,构建犬瘟热N基因原核表达重组质粒,然后转化大肠杆菌BL21(DE3)菌株,经IPTG诱导表达重组N蛋白。从包涵体中纯化重组蛋白,制备多克隆抗体,采用Western blot检测其特异性。结果显示PCR扩增得到犬瘟热N基因,重组N蛋白在大肠杆菌BL21(DE3)菌株中得到表达,制备的多克隆抗体能与N蛋白特异性反应。To study the function of N protein of Canine distemper virns （CDV）, the CDV-N gene was amplified in PCR and cloned into expression vector pET-32a（＋）. Subsequently, the recombinant plasmid pET-32a-N was transformed into E.coli BL21 （DE3） cells prior to induction with IPTG. The CDV-N protein was purified from the inclusion bodies. The PCR product of the CDV N gene was 1572 bp in length. The recombinant N protein was used to immunize rabbits to produce polyclonal antibodies. The rabbit antiserum was sampled and its specificity for N protein was confirmed in Western blot.

关 键 词：犬瘟热病毒 N蛋白 原核表达 多克隆抗体 

分 类 号：S852.659.5[农业科学—基础兽医学]