中文摘要：

目的在真核细胞中表达人磷脂氢谷胱甘肽过氧化物酶（PHGPx）并制备抗PHGPx多克隆抗体。方法用特异引物从人睾丸组织cDNA文库中扩增出人线粒体型PHGPx cDNA，与真核表达载体pcDNA3，1／His重组，在COS．1细胞内表达PHGPx。用大肠杆菌表达的突变的PHGPx免疫动物，制备抗PHGPx抗体。结果重组体pcDNA-PHGPx转染的COS-1细胞表达了人PHGPx融合蛋白，PHGPx活性为111．5μmol·mg^-1·min^-1。高于对照组5倍。免疫扩散测得抗PHGPx多克隆抗血清滴度为1：16；用于蛋白印迹时稀释倍数为1：500。可见特异免疫沉淀条带。结论人PHGPx在COS-1细胞内获得了表达．抗PHGPx多克隆抗体有特异性。可以用于重组表达的PHGPx的鉴定。

英文摘要：

Objective To express human phospholipid hydroperoxide glutathione peroxidase （PHGPx） in eukaryotic cells and to prepare its polyclonal antibody. Methods PHGPx cDNA was amplified from a human testis library using specific primers and cloned into expression vector pcDNA3.1/His. Expression of PHGPx was performed in COS-I cells. Polyclonal antibody against PHGPx was prepared using mutated PHGPx expressed in E.coli. Results The activity of the PHGPx fusion protein expressed in COS-1 cells transfected with recombinant pcDNA-PHGPx was 111.5 μmol·mg^-1·min^-1 ,5-fold higher than that of control group. The titer of anit-PHGPx polyclonal antibody was 1 ： 16 obtained by immunoprecipitation assay,and it could be diluted into 1 ： 500 when used in Western Blotting. The specificity of the polyclonal antibody was verified by the specific immunoprecipitation straps. Conclusions Recombinant PHGPx is expressed in COS-I cells and anit-PHGPx antibody can be used to identify the recombinant PHGPx.