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目的在大肠杆菌中表达富含组氨酸蛋白Ⅱ，为抗疟疫苗研究及制备疟疾诊断单克隆抗体提供实验材料。方法采用ＰＣＲ方法特异性扩增恶性疟原虫云南株（ＰＦＤ—３／ＹＮ）富含组氨酸蛋白Ⅱ部分基因，经基因序列测定后克隆于ｐＧＥＸ４Ｔ－１融合蛋白的表达，并用ｄｏ－ＥＬＩＳＡ对表达产物进行鉴定。结果ＨＲＰⅡ基因与ｐＧＥＸ４Ｔ－１重组后在大肠杆菌ＴＧ１中表达—４１ｋＤａ的融合蛋白。Ｄｏｔ－ＥＬＩＳＡ鉴定表明ＨＲＰⅡ基因表达产物能被抗ＨＲＰⅡ单克隆抗体所识别。结论大肠杆菌表达的ＨＲＰⅡ能与抗ＨＲＰⅡ单克隆抗体发生反应，且具有恶性疟原虫ＨＲＰⅡ抗原表位。
Objective To express histidine-rich protein Ⅱ in Escherichia coli and provide experimental materials for the research of malaria vaccine and preparation of monoclonal antibody for diagnosis of malaria. Methods Partial amplification of the histidine-rich protein Ⅱ gene of Plasmodium falciparum (PFD-3 / YN) by PCR was performed. The gene was cloned into pGEX4T-1 fusion protein and sequenced by do-ELISA The expression product was identified. Results After recombination of HRPⅡ gene and pGEX4T-1, a -41kDa fusion protein was expressed in E. coli TG1. Dot-ELISA identification showed that the HRPII gene product was recognized by anti-HRPII monoclonal antibody. Conclusion The HRP Ⅱ expressed in E. coli can react with anti-HRPⅡ monoclonal antibody and has the antigen of Plasmodium falciparum HRPⅡ.