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目的：构建编码恶性疟原虫复合多价保护性抗原的基因的重组质粒，为进行基因免疫提供条件。方法：设计一对特异引物Ｐ１与Ｐ２，采用ＰＣＲ法扩增获取ＭＳＰ１中Ｃ端１９肽基因，纯化后用ＳａｌⅠ＋ＸｂａⅠ双酶切，把含复合基因ＰｆＣＭＲ的重组质粒ｐＷＲ４５０－１／ＰｆＣＭＲ用ＥｃｏＲⅠ＋ＳａｌⅠ双酶切，回收复合基因ＰｆＣＭＲ，将ＭＳＰ１１９基因与ＰｆＣＭＲ基因串联后与经ＥｃｏＲⅠ＋ＸｂａⅠ双酶切的真核表达质粒ｐｃＤＮＡ３进行重组，转化大肠杆菌ＪＭ１０９，经电泳初筛、双酶切鉴定及ＰＣＲ鉴定。结果：ＰＣＲ扩增获得３６３ｂｐ的ＭＳＰ１１９基因，重组克隆经双酶切鉴定及ＰＣＲ鉴定后获得正确重组克隆子ｐｃＤＮＡ３－ＰｆＣＭＲ－ＭＳＰ１１９（命名为ｐｃＤＮＡ３－Ｐｆ８），Ｐｆ８基因长度为６１８ｂｐ。结论：成功构建编码恶性疟原虫多价保护性抗原的基因的重组质粒ｐｃＤＮＡ３－Ｐｆ８。
OBJECTIVE: To construct a recombinant plasmid encoding the gene encoding multivalent protective antigen of Plasmodium falciparum, which provides conditions for gene immunization. Methods: A pair of specific primers P1 and P2 were designed and synthesized. The C-terminal 19 peptide of MSP1 was amplified by PCR and purified with SalⅠ + XbaⅠ. The recombinant plasmid pWR450-1 / PfCMR containing PfCMR was digested with EcoRⅠ + The gene PfCMR was cut and recovered. The MSP119 gene and PfCMR gene were ligated with the recombinant plasmid pcDNA3, which was digested with EcoRⅠ + XbaⅠ. The recombinant plasmid was transformed into Escherichia coli JM109 and identified by restriction enzyme digestion and PCR. Results: The 363bp MSP119 gene was obtained by PCR amplification. The recombinant clone pcDNA3-PfCMR-MSP119 (named as pcDNA3-Pf8) was double-digested and identified by PCR. The length of Pf8 gene was 618bp. Conclusion: The recombinant plasmid pcDNA3-Pf8 encoding the gene of multivalent protective antigen of Plasmodium falciparum was successfully constructed.