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.
出处
《中国动物传染病学报》
CAS
北大核心
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)
关键词
鸡β干扰素
原核表达
抗病毒活性
Chicken interferon-β
prokaryotic expression
antiviral effect
