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目的构建恶性疟原虫FCC—1／HN株红细胞结合抗原（EBA—175）和富组氨酸蛋白Ⅱ（HRP─Ⅱ）抗原的重组质粒并进行初步鉴定。方法通过聚合酶链反应（PCR）对恶性疟原虫FCC—1／HN株的EBA─175和HRP─Ⅱ基因进行体外扩增，用HindⅢ／BamHI和BamHI／EcoRI分别消化扩增产物，定向克隆pcDNA3载体，转化至感受态大肠杆菌TG1，经抗性筛选和快速凝胶电泳鉴定，再经PCR和酶切鉴定。结果从恶性疟原虫FCC—1／HN株基因组DNA中扩增出EBA—175和HRP─Ⅱ两基因片段并定向插入PCDNA3载体构建重组质粒。结论所获重组克隆含有编码恶性疟原虫FCC—1／HN株EBA—175和HRP—Ⅱ的基因片断。
Objective To construct recombinant plasmids of Plasmodium falciparum FCC-1 / HN erythrocyte binding antigen (EBA-175) and histidine-rich protein Ⅱ (HRP-Ⅱ) antigen and preliminary identification. Methods The EBA175 and HRPⅡ genes of Plasmodium falciparum FCC-1 / HN strain were amplified by polymerase chain reaction (PCR) in vitro. The amplified product was digested with HindⅢ / BamHI and BamHI / EcoRI, respectively. The recombinant plasmid pcDNA3 The vector was transformed into competent E. coli TG1, identified by resistance screening and rapid gel electrophoresis, and then identified by PCR and restriction enzyme digestion. Results Two fragments of EBA-175 and HRP-Ⅱ were amplified from genomic DNA of Plasmodium falciparum FCC-1 / HN strain and inserted into PCDNA3 vector to construct a recombinant plasmid. Conclusion The obtained recombinant clone contains the gene fragment encoding the EBA-175 and HRP-Ⅱ of Plasmodium falciparum FCC-1 / HN strain.