【摘要】 目的　研究人Lipocalin型前列腺素D合酶 (L PGDS)的编码蛋白质在大肠杆菌中的表达 ,进一步制备抗L PGDS编码蛋白质的多克隆抗体。方法　用逆转录多聚酶链式反应 (RT PCR)方法克隆L PGDS基因蛋白编码序列。构建该基因的表达质粒 ,在大肠杆菌中诱导表达 ,SDS PAGE分析表达的蛋白质。用RediPackGST亲和层析柱纯化的蛋白质免疫BALB/c小鼠制备多抗。结果　RT PCR所分离的L PGDS基因蛋白编码序列 ,经测序证明与基因库所报道的L PGDS的基因序列完全一致。SDS PAGE分析证实该基因编码的蛋白质在大肠杆菌中表达 ,并获得效价为 1∶6 40 0的多克隆抗体。结论　L PGDS基因编码蛋白质能够在原核细胞中表达 ,并能获得抗L PGDS基因编码蛋白质的多克隆抗体 ,为进一步深入研究该蛋白质的结构与功能打下了基础更多还原

【Abstract】 Purpose To express lipocalin prostaglandin D synthase gene(L?PGDS) in E.coli and prepare anti LPGDS polyclonal antibody. Methods The human L?PGDS was cloned by RT?PCR.The expression plasmid of pGEX4T?1?LPGDS was constructed and transformed into E.coli BL21.The fusion protein was induced to express by IPTG and analyzed by SDS?PAGE.BALB/c mice were immunized with the protein purified by RediPack GST Purification Modules for the preparation of polyclonal antibody. Results Sequence analysis of gene of L?PGDS revealed identity to the L?PGDS reported.Expression of the protein in E.coli was verified by SDS?PAGE.The polyclonal antibody obtained from immunized BALB/c mice had a title of 1∶6 400. Conclusions Purified protein and antibody is useful for further study of structure and function of the L?PGDS.更多还原