绵羊特异新基因OaN2片段的克隆及融合表达  

Cloning and Fusion Expression of a Novel Ovine Gene Fragment

作　　者：李刚[1] 刘卡[2] 高剑峰[1] 马润林[2] 

机构地区：[1]石河子大学生命科学学院,石河子832000 [2]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《石河子大学学报（自然科学版）》2010年第1期43-46,共4页Journal of Shihezi University(Natural Science)

摘　　要：为制备绵羊MHC区段1个新基因(OaN2)的特异性抗体及其表达模式和功能研究奠定基础,克隆OaN2的1个特殊片段(OaN2F,Genbank登录号:GQ244478)并做融合表达。以反转录得到的中国美利奴细毛羊的第一链cDNA为模板进行PCR扩增,并利用扩增产物构建重组质粒,转化感受态大肠杆菌DH5α后诱导表达得到OaN2F和GST(谷胱甘肽S转移酶)的融合蛋白GST-OaN2F,将其纯化后鉴定。结果表明,成功克隆到了OaN2F(111bp)片段,并得到了纯化的30kD的GST-OaN2F融合蛋白。A novel gene fragment（OaN2F, Genbank accession number： GQ244478） of MHC region in ovine was cloned and fusion expression was performed in order to obtain its specific antibody and provide a basis for further study on expression profile and function. The first strand cDNA was synthesized by reverse transcription PCR in Chinese fine-wool Merino sheep as template was amplified by PCR. The products of PCR was constructed the recombinant plasmid and induced with IPTG for expression of fusion protein. Fusion protein with glutathione S-transferase tag could be purified by glutathione-agarose affinity Chromatography. Purified protein was identified by SDS-PAGE. The result shows the cDNA of OaN2F was successfully cloned,and the length of cDNA fragment was lllbp. The fusion protein GST-OaN2F（30kD） was expressed and purified. The conclusion is that the cloning of OaN2F and the expression of GST-OaN2F provide a basis for preparation of specific antibody .

关 键 词：绵羊 MHC OaN2F 融合表达 

分 类 号：S826[农业科学—畜牧学] Q78[农业科学—畜牧兽医]