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.
出处
《中国动物传染病学报》
CAS
北大核心
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)
关键词
犬瘟热病毒
N蛋白
原核表达
多克隆抗体
Canine distemper virns （CDV）
N protein
prokaryotic expression
polyclonal antibody
