鸡IFN-γ基因的原核表达及其表达产物的抗NDV活性  

Prokaryotic expression of chicken interferon-γ gene and detection of activity of the expressed product against NDV

作　　者：董建宝[1] 黄溢泓[2] 谢芝勋[1] 孙建华[1] 刘加波[1] 庞耀珊[1] 邓显文[1] 谢丽基[1] 

机构地区：[1]广西兽医研究所,广西南宁530001 [2]广西柳州市动物疫病预防控制中心,广西柳州545005

出　　处：《中国兽医科学》2007年第10期887-891,共5页Chinese Veterinary Science

基　　金：广西留学基金项目(桂科回0542006)

摘　　要：设计了1对特异性引物,从含有广西三黄鸡IFN-γ基因的重组质粒pMD18-ChIFN-γ中扩增出鸡的IFN-γ基因,将该基因插入到原核表达载体pGEX-4T-1中构建成重组表达质粒pGEX-ChIFN-γ,并将其转入到大肠杆菌BL21中,IPTG诱导表达的融合蛋白经SDS-PAGE和Western-blot分析,其分子质量约为42.4 ku,表明鸡IFN-γ基因在原核细胞中得到了表达。采用NDV国内参考强毒株F48E9对复性后的IFN-γ进行了活性测定;结果显示,IFN-γ的抗病毒活性为1.33×104U/mL。One pair of specific primers was designed to amplify an interferon-γ gene from the recombinant plasmid pMD18-ChIFN-γ. Then the interferon-γ gene was inserted into prokaryotic expression vector pGEX-4T-1. The recombinant vector pGEX-ChIFN-γ was transformed into Escherichia coli BL21 competent cells and induced by IPTG. Analyses of SDS-PAGE and Western-blotting showed that a protein band was produced with a molecular mass of approximate 42.4 ku,indicating that the protein of ChIFN-γ gene was correctly expressed in the transformed bacteria. Newcastle disease virus F48E9 strain was used to detect the bioactivity of renatured ChIFN-γ,and the results showed that the bioactivity was 1.33 × 10^4 U/mL.

关 键 词：鸡 Γ-干扰素基因 原核表达 抗病毒活性 

分 类 号：S859.797[农业科学—临床兽医学]