小鼠IL-12受体β2胞内区蛋白表达及多抗制备  

Expression of Cytoplasmic Domain of Mouse Interleukin 12 Receptor β2 and Preparation of Polyclonal Antibodies

作　　者：林瑛[1,2] 陈婉蓉[1] 季永镛[2] 孙兵[2] 

机构地区：[1]华东师范大学生命科学学院,上海200062 [2]中国科学院上海生物化学与细胞生物学研究所,上海200031

出　　处：《上海免疫学杂志》2003年第5期293-297,共5页Shanghai Journal of Immunology

摘　　要：从ConA刺激的小鼠脾细胞中抽提总RNA,用RT-PCR的方法克隆出小鼠IL-12R β2亚基胞内区基因。再将其插入到原核表达载体pGEX-4T-1中,转化宿主菌BL-21(DE3),用IPTC诱导宿主菌表达蛋白。SDS-PAGE分析结果表明,蛋白主要存在于包涵体中。利用割胶回收的方法纯化目的蛋白,免疫兔子制备多抗。经ELISA检测,证明免疫后兔血清中有高滴度的针对IL-12Rβ2亚基胞内区的抗体。The gene of cytoplasmic domain of mouse interleukin 12 receptor (32 was amplified from ConA stimulated mouse spleen cells via RT-PCR. It was cloned into the prokaryotic expression vector pGEX-4T-1. Then the recombinant plasmid was transformed into E. coli BL-21 (DE3 ) . The fusion protein of GST-mIL-12R (32 was expressed under the condition of IPTG induction, and was found to be located in inclusion bodies. The protein was purified from SDS-PAGE gel and was used to immunize rabbit to produce polyclonal antibodies against murine IL-12R (32. When the serum antibody was measured by ELISA, it was demonstrated that the rabbit serum antibody could specifically recognize cytoplasmic domain of murine interleukin 12 receptor β2.

关 键 词：IL-12Rβ2 基因克隆 蛋白表达 多克隆抗体 

分 类 号：R392.11[医药卫生—免疫学]