hPLIC-2可溶性表达及多克隆抗体的制备  

Expression of hPLIC-2 and antibody preparation

作　　者：李楚芳[1] 刘萱[1] 李平[1] 马清钧[1] 曹诚军 

机构地区：[1]事医学科学院生物工程研究所,北京100850

出　　处：《生物技术通讯》2004年第5期451-453,共3页Letters in Biotechnology

摘　　要：利用多聚酶联反应(PCR)扩增获得人hPLIC-2基因,并将该基因克隆到原核表达载体pGEX-2T上。转化大肠杆菌JM109,在27℃经IPTG诱导后,GST-hPLIC-2融合蛋白获得高效可溶性表达,并通过GlutathioneSephorase4B获得纯化。使用结合有GST-hPLIC-2的GlutathioneSephorase4B凝胶珠免疫大白兔制备了兔抗hPLIC-2多克隆抗体,抗体滴度大于1∶16000。该抗体制备为研究hPLIC-2的功能提供了有用的工具。hPLIC-2 gene was amplified by PCR and inserted into pGEX-2T vector. The recombinant plasmid was transformed into host cell JM109. GST-hPLIC-2 fusion protein was expressed at a high level as a soluble form after being induced by IPTG at 27 ℃. The soluble fusion protein was purified by Glutathione Sephorase 4B. The polyclonal antibody was made after immunizing rabbit by Glutathione Sephorase 4B binding GST-hPLIC-2 and the titer of the antiserum was more than 1∶16 000. This antibody will play important role in the functional study of hPLIC-2 gene.

关 键 词：多克隆抗体 GST 可溶性表达 多聚酶 大白兔 抗体滴度 抗体制备 IPTG PCR) 原核表达载体 

分 类 号：Q786[生物学—分子生物学] R392[医药卫生—免疫学]