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.
出处
《生物技术通讯》
CAS
2004年第5期451-453,共3页
Letters in Biotechnology
