【摘要】 目的　在 Ｈｅ Ｌａ 细胞中表达恶性疟原虫红细胞结合蛋白 Ｅ Ｂ Ａ－ １７５ 和富组氨酸蛋白 Ｈ Ｒ Ｐ－ Ｉ Ｉ， 进一步研究其生物学功能和免疫学特性。方法　将 Ｅ Ｂ Ａ－ １７５ 和 Ｈ Ｒ Ｐ－ Ｉ Ｉ 部分基因串接插入 Ｐ Ｃ Ｄ Ｎ Ａ３ 真核表达载体， 获得重组表达质粒 Ｐ Ｃ Ｄ Ｎ Ａ３ Ｅ Ｂ Ａ１７５ － Ｈ Ｒ Ｐ Ｉ Ｉ。采用磷酸钙－ Ｄ Ｎ Ａ 共沉淀法将重组质粒转染 Ｈｅ Ｌａ 细胞进行体外表达， 对表达产物进行 Ｓ Ｄ Ｓ－ Ｐ Ａ Ｇ Ｅ、 Ｄｏｔ－ Ｅ Ｌ Ｉ Ｓ Ａ 和 Ｗｅｓｔｅｒｎ － ｂｌｏｔ 鉴定。结果　表达产物占表达蛋白总量的１６４ ％ ， 相对分子量与预设计的４８ｋ Ｄａ 基本相符。表达产物与鼠抗恶性疟原虫血清及构建的重组质粒免疫鼠血清产生特异免疫反应。结论　表达载体 Ｐ Ｃ Ｄ Ｎ Ａ３ 在 Ｈｅ Ｌａ 细胞内可表达出具有一定免疫学活性的恶性疟原虫重组抗原蛋白。更多还原

【Abstract】 AimTo express erythrocyte binding antigen 175 and histidine rich protein II of P falciparum in HeLa cell line and to further study their biological and immunological functions.Methods Two genes encoding for EBA-175 and HRP-II were directly cloned into eukaryotic expression vector pCDNA3.The constructed recombinant plasmid PCDNA3/EBA175-HRPII was transfected into HeLa cell line by the calcium phosphobe priciptation employing Calcium precipitation method.The HeLa cells were cultured in 1640 selective medium containing G418 to screen positive cell clones which could expressed stablely EBA-175 and HRP-II.The expressed protein was isolated and analysed by SDS-PAGE,Dot-ELISA and Western-blot assays.Results The analysis of SDS-PAGE indicated that the molecular weight of the expressed recombinant protein was about 48kDa and amounted to 16 4% of the total proteins in the HeLa cells.The expressed potein could react specifically with the sera from the mice immunized with P falciparum and those with recombinant plasmid DNA by Dot-ELISA and Western-blot.ConclusionHeLa cell line transfected with recombinant plasmid pCDNA3/EBA175-HRPII could express the right antigenic protein of P falciparum in vitro.The expressed protein had certain immonological activity which may bring about the possibility of malaria vaccine.更多还原