重组人卵透明带蛋白3的制备及其免疫学活性分析

Preparation of recombinant human zona pellucida 3 protein and its immunologic activity

目的:纯化制备rhuZP3,并分析其免疫学活性。方法:在含重组质粒pGEX4T-1/huZP3的大肠杆菌BL21中,经IPTG诱导表达出GST-融合蛋白,蛋白经过一系列的纯化,然后SDS-PAGE电泳鉴定蛋白纯度。rhuZP3免疫小鼠,ELISA法检测抗血清对rhZP3的抗体反应。结果:表达出了可溶性融合蛋白,纯化后的rhuZP3纯度达95%。而且它在ELISA鉴定实验中能被抗rhuZP3抗体识别。结论:通过原核表达系统制备的rhuZP3及其抗体具有免疫学活性。

Objective： To prepare and purify recombinant human zona pellucida 3 protein, and to study its immunologic activity. Methods：The E. coli BL21 containing recombinant plasmid pGEX4T-1/huZP3 was induced to express GST-fusion protein by IPTG. After a series of purification procedure, the purified protein was analyzed by SDS-PAGE. After the mice immunized with rhuZP3, the antibody responses against rhuZP3 were detected by ELISA. Results： The soluble fusion protein was expressed,and purity of rhuZP3 was 95%. Moreover, purity of rhuZP3 could be recognized by anti-human ZP3 in ELISA. Conclusion：The rhuZP3 is obtained through the preparation of prokaryotic expression system and anti-rhuZP3 antibody has immunological activity.