鸡β干扰素的原核表达及抗病毒活性检测  

PROKARYOTIC EXPRESSION AND ANTIVIRAL EFFECT OF CHICKEN INTERFERON-β

作　　者：吴斯宇 黄健妮 崔进[1] 冯赛祥[1] 欧阳国文[1] 向程威 郭旸[1] 廖明[1] 焦培荣[1] 

机构地区：[1]华南农业大学兽医学院人兽共患病防控制剂国家地方联合工程实验室,广州510642

出　　处：《中国动物传染病学报》2016年第4期59-63,共5页Chinese Journal of Animal Infectious Diseases

基　　金：科技部国际合作项目(2013DFA31940);广州市科技计划项目(2013J4500030;201300000037)

摘　　要：根据大肠杆菌密码子的偏好性对GenBank中发表的鸡β干扰素基因序列进行密码子优化与人工合成,构建原核表达质粒pET-28b-ChIFN-β,重组表达质粒转化大肠杆菌BL21（DE3）,经IPTG诱导表达,产物主要以包涵体形式存在。包涵体进行变性、复性和镍柱亲和纯化后,SDS-PAGE、Western blot分析表明pET-28b-ChIFN-β表达正确,并且表达量较高。重组鸡β干扰素蛋白在DF-1细胞上能明显抑制水疱性口炎病毒繁殖,抗病毒活性达2.73×10-6UI/mg。本研究结果为鸡β干扰素在临床防治禽类病毒性疾病的探索奠定了基础。Chicken interferon-p（ChIFN-P） gene sequence from GenBank was optimized and synthesized based on the codon preference of E.coli.The ChIFN-P gene was cloned into prokaryotic expression vector pET-28 b.Analysis of SDS-PAGE and Western blot indicated that pET-28b-ChIFN-P was correctly and highly expressed in E.coli induced with IPTG.The expressed protein was purified via nickel affinity chromatography after being denatured and re-natured.The purified recombinant ChIFN-P protein was able to inhibit vesicular stomatitis virus replication at 2.73× 10-6 UI/mg on DF-1 cells.The results of the present study would be benefit to the research of chicken interferon β regarding prevention and treatment of avian viral diseases.

关 键 词：鸡β干扰素 原核表达 抗病毒活性 

分 类 号：S852.42[农业科学—基础兽医学]