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鸡IFN-γ基因的原核表达及其表达产物的
抗NDV活性
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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
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（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）
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摘要：
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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42.4ku，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。 |
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关键词：
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鸡；γ-干扰素基因；原核表达；抗病毒活性 |
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中图分类号：S 859.797 文献标识码：
A 文章编号：1673-4696(2007)10-0887-05 |
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Prokaryotic expression of chicken interferon-γ gene and detection of
activity of the expressed product against NDV |
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DONG Jian-bao1,HUANG Yi-hong2,XIE
Zhi-xun1,SUN Jian-hua1,LIU Jia-bo1,
PANG Yao-shan1,DENG Xian-wen1,XIE Li-ji1 |
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(1.Guangxi Veterinary Research Institute,Nanning 530001,China;
2.Liuzhou Prefecture Center for Animal Disease Control and Prevention,Liuzhou 545005,China) |
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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 42.4ku,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. |
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Key words: chicken;interferon-γ gene;prokaryotic expression;antiviral activity |
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