【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。更多还原

【Abstract】 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 1.33×104U/mL.更多还原