北极狐γ-干扰素cDNA的克隆、表达及活性测定  

Cloning,Expression and Antiviral Activity of Arctic Fox (Alopex lagopus) Interferon-γ Gene

作　　者：张海玲[1] 柴秀丽[1] 罗国良[1] 王凤雪[1] 易立[1] 邵西群[1] 闫喜军[1] 

机构地区：[1]中国农业科学院特产研究所,吉林132109

出　　处：《生物工程学报》2008年第9期1625-1630,共6页Chinese Journal of Biotechnology

基　　金：吉林省科技发展计划项目(No.20060550)资助~~

摘　　要：为研究狐γ-干扰素的生物学活性,应用反转录聚合酶链式反应(RT-PCR)从北极狐外周血淋巴细胞中扩增出γ-干扰素(VuIFN-γ)cDNA。序列分析表明VuIFN-γcDNA全长501bp,编码23个氨基酸的信号肽和144个氨基酸的成熟肽蛋白,与已发表的银黑狐和犬IFN-γ核苷酸序列同源性为99.8%和99.4%;氨基酸同源性均为100%。应用原核表达系统高效表达北极狐γ-干扰素成熟肽蛋白,SDS-PAGE和Western blotting分析表达的融合蛋白分子量约为19kD,以不溶性的包涵体形式存在。重组蛋白经纯化和复性,在Vero和MDCK细胞上可明显抑制VSV病毒的复制,并测出北极狐重组γ-干扰素的活性单位分别为1.0×106u/mg和1.56×105u/mg,为进一步开发基因工程狐干扰素奠定基础。In order to characterize the biological activity of fox （Vulpes vulpes） interferon gamma（VuIFN-γ）, We have isolated the cDNA encoding arctic fox（Alopex lagopus）VuIFN-γ. This cDNA encodes a 23 amino acid signal peptide and a 144 amino acid mature protein, which shares 99.8% or 99.4% for nucleotide identity with silver fox and canine, respectively, and 100% for amino acid identity. Expression of recombinant mature arctic fox interferon gamma （mVuIFN-γ） in bacterial system was confirmed by SDS-PAGE and Western blotting analysis. Recombinant VuIFN-γ showed higher antiviral activity against vesicular stomatitis virus in cultured Vero and MDCK by inhibiting vires induced cytopathic effect, In view of the immunomodulatory and antiviral activities of VuIFN-γ, it may provide a basis for further research on antiviral therapy of recombinant VuIFN-γ in economic animal practice.

关 键 词：北极狐 干扰素-Γ 抗病毒活性 

分 类 号：Q78[生物学—分子生物学] Q785