猪SIRT1基因的克隆、表达及多克隆抗体制备  

Cloning,expression and preparation of polyclonal antibody of porcine SIRT1 gene

作　　者：江学斌[1] 马苗鹏[2] 肖淑君[2] 司徒嘉欣 杨军[2] 施巨清 蔡海明[2] 张玲华[2] 

机构地区：[1]广州大学生物工程研究所,广州510006 [2]华南农业大学生命科学学院/广东省农业生物蛋白质功能与调控重点实验室,广州510642

出　　处：《华中农业大学学报》2014年第5期80-84,共5页Journal of Huazhong Agricultural University

基　　金：广东省农业攻关项目(2012B020305007);广东省教育部产学研结合项目(2011B090400471)

摘　　要：根据GenBank中猪SIRT1mRNA序列设计引物,使用杜大长商品猪大脑RNA经RT-PCR扩增得到猪SIRT1基因全长表达序列,通过Ω-PCR将SIRT1蛋白末端抗原表位序列插入载体pET-28a(+),在大肠杆菌中表达并纯化得到45ku大小的蛋白,以其免疫新西兰大白兔并制备出抗体滴度达212的多克隆抗体。结果表明,成功制备出可以特异性识别猪SIRT1蛋白的抗体。Based on the porcine SIRT1 mRNA sequence from GenBank,primers of porcine SIRT1 gene were designed and coding region of SIRT1 gene was amplified.The epitope sequences located on the rear section of SIRT1 protein were then inserted into pET-28a(+)byΩ-PCR.The SIRT1 protein was expressed in E.coli,purified and its polyclonal antibody was produced by immuning rabbit,the protein weight is about 45ku and the antibody titers is about 212.The results showed that antibody is capable of detecting the porcine SIRT1 protein specifically and provided a new tool for studies of the cellular path way involving in porcine SIRT1.

关 键 词：猪 SIRT1 PCR 多克隆抗体 

分 类 号：Q786[生物学—分子生物学] S828[农业科学—畜牧学]