重组犬IFN-γ在HEK293T细胞中的表达  

Expression of recombinant canine IFN-γ gene in HEK 293T cells

作　　者：靳慧君[1] 李楠[1] 郭常春 韩冬梅[1] 王微[1] 仲飞[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001 [2]广州天王动物保健品有限公司,广东广州510800

出　　处：《河北农业大学学报》2009年第2期102-105,共4页Journal of Hebei Agricultural University

基　　金：国家自然科学基金资助(30771586)

摘　　要：为了建立一套犬IFN-γ(cIFN-γ)在HEK293T细胞中表达的方法,用伴刀豆球蛋白A(ConA)刺激犬脾细胞,通过RT-PCR方法从脾细胞中扩增犬IFN-γ基因,并将其克隆到pcDNA3.1A载体中,构建的真核表达载体pcDNA3.1A-cIFN-γ经磷酸钙介导转染HEK293T细胞进行表达。结果表明:克隆的犬IFN-γcDNA基因与GenBank上发表的序列一致,同源性100%。表达产物经Western-blot方法检测,证明克隆的犬IFN-γ基因能够在HEK293T细胞中进行表达,并且表达产物能够分泌到细胞外。To establish the method of canine IFN-γ （cIFN-γ） expression in HEK293T cells, the canine spleen cells were stimulated by concanavalin A. The cDNA encoding cIFN-γ was amplified by RT- PCR from the stimulated cells. Then the cIFN-γ gene was inserted into eukaryotic ex- pression vector pcDNA3.1A. The recombinant pcDNA3.1A - cIFN-γ plasmids were transfected into HEK293T cells mediated by calcium phosphate. Results showed that the sequence of cIFN-γ cDNA amplified was identical to that published in GenBank, and the homology is 100 %. The expressed products were detected by Western - blot, which indicated that cIFN -γ gene could be ex pressed in HEK293T cells and secreted from the cells.

关 键 词：犬γ-干扰素 HEK293T细胞 表达 

分 类 号：S829.2[农业科学—畜牧学]