细胞周期蛋白质依赖性激酶CDK2多克隆抗体的制备和鉴定  

Preparation and identification of the polyclonal antibodies against cyclin-dependent kinase 2

作　　者：李清[1] 陈越[1] 陈勇[1] 佟立全[1] 孔维[1] 金英花[1] 

机构地区：[1]吉林大学生命科学学院

出　　处：《辽宁师范大学学报（自然科学版）》2007年第4期491-494,共4页Journal of Liaoning Normal University:Natural Science Edition

基　　金：国家自然科学基金委主任基金资助项目(30640064)

摘　　要：我们采用PCR技术合成编码CDK2肽段的基因,将其置于谷胱甘肽转移酶(GST)编码基因的下游,在IPTG诱导下,于E.coli中诱导表达了GST-CDK2肽融合蛋白质,以此融合蛋白质作为免疫原免疫家兔制备抗CDK2的多克隆抗体.经Western Blot检测证明:该抗体能够特异地识别CDK2蛋白质,可作为CDK2的特异性检测抗体,用于研究细胞周期和细胞凋亡进程中CDK2的作用.The gene coding CDK2 specific peptide were synthesized with PCR and cloned into C-terminal of GST expression vector pGEX-6P-1. The fusion proteins, GST-CDK2 peptide, were expressed in E. coli（BL21） by adding 0.1 mmol/L IPTG and purified with Glutathione Sepharose 4B. The rabbit was immunized with the mixtures of purified fusion proteins and complete Freund＇s adjuvants. The sera of immunized rabbit were obtained and then analyzed by Western Blot. The result showed that these polyclonal antibodies in the sera can recognize the human CDK2 proteins from HeLa cells. These data suggested that the sera can be used as specific antibodies against CDK2 proteins in studying the role of CDK2 in the process of cell cycle and apoptosis.

关 键 词：CDK2 多克隆抗体 WESTERN BLOT 

分 类 号：Q789[生物学—分子生物学]