远缘链球菌细胞分裂蛋白的表达、纯化和抗体的制备  

Expression of streptococcus sobrinus's fts Kprotein and preparation of polyclonal antibody

作　　者：金涛[1] 吴补领[1] 苏凌云[1] 高杰[1] 

机构地区：[1]第四军医大学口腔医学院,西安710032

出　　处：《陕西医学杂志》2005年第5期518-520,共3页Shaanxi Medical Journal

基　　金：第四军医大学创新工程资助项目(NoCX02A012)

摘　　要：目的表达与纯化远缘链球菌克隆细胞分裂蛋白(ftsK),制备多克隆抗体并鉴定。方法利用IPTG诱导ftsK在大肠杆菌中表达,随后以再生Ni-NTA柱纯化蛋白。纯化后的蛋白多次注射于新西兰大白兔中,获取多克隆抗血清,ELISA法检测抗体效价,Westernblot法检测抗体特异性。结果获得了纯度较高的远缘链球菌ftsK蛋白及其抗体,多抗效价达到1∶10000,特异性较好。结论成功表达了远缘链球菌ftsK蛋白,获得了效价较高、特异性较强的多克隆抗体,为进一步研究ftsK蛋白功能获取了良好的工具。Objective: To express the streptococcus sobrinus's fts K protein and prepare polyclonal antibody against the protein. Methods: IPTG was used to induce the expression of fts K protein, Ni-NTA column was used to purify the protein. Polyclonal antibody was prepared by immunizing rabbits with this protein. The expressed protein was identified by ELISA and western blot. Results: The purified streptococcus sobrinus's fts K protein was obtained, the title of prepared polyclonal antibody is 1∶10000, and the antibody's specificity is very high. Conclusion: The purified streptococcus sobrinus's fts K protein and polyclonal antibody was successfully obtained. The expression of fts K in E coli provided the basic material for studying the role of fts K, and polyclonal antibody is a good tool for the function research of fts K.

关 键 词：远缘链球菌 细胞分裂 WESTERNBLOT法 制备 ELISA法检测 多克隆抗体 新西兰大白兔 多克隆抗血清 表达与纯化 抗体特异性 大肠杆菌 纯化蛋白 多次注射 抗体效价 蛋白功能 NTA 步研究 

分 类 号：R780.2[医药卫生—口腔医学] R735.7[医药卫生—临床医学]