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目的 构建恶性疟原虫组氨酸富集蛋白Ⅱ (Histidine richproteinⅡ ,HRPⅡ )反义真核表达载体 ,为研究HRPⅡ反义核酸的抗疟作用奠定基础。方法 应用基因重组技术 ,将我们已经克隆的恶性疟原虫HRPⅡ基因从HRPⅡ /pUC19反向亚克隆入荧光真核表达载体pEGFP N3,用含卡那霉素LB培养基平板筛选转化菌 ,阳性重组子经HindⅢ和SacⅠ双酶切分析、PCR鉴定。结果 恶性疟原虫HRPⅡ基因从HRPⅡ /pUC19重组质粒中 ,成功地亚克隆至真核表达载体pEGFP N3,获得了反义HRPⅡ /pUC19重组质粒。结论 恶性疟原虫HRPⅡ基因以反方向亚克隆入pEGFP N3质粒 ,成功构建反义HRPⅡ /pEGFP N3荧光真核表达载体 ,解决了HRPⅡ反义核酸来源问题 ,为下一步研究反义HPRⅡ的抗疟作用奠定了基础。
Objective To construct an antisense eukaryotic expression vector for Histidine rich protein Ⅱ (HRP Ⅱ) of Plasmodium falciparum, and lay the foundation for the study of antimalarial activity of HRP Ⅱ antisense nucleic acid. Methods The gene of recombinant Plasmodium falciparum HRPⅡ was cloned from HRP Ⅱ / pUC19 into the eukaryotic expression vector pEGFP N3. The recombinant plasmids were used to screen transformed cells with kanamycin LB medium. The positive recombinant The Hind Ⅲ and Sac Ⅰ digestion analysis, PCR identification. Results Plasmodium falciparum HRPⅡ gene was successfully subcloned into eukaryotic expression vector pEGFP N3 from HRPⅡ / pUC19 recombinant plasmid, and antisense HRPⅡ / pUC19 recombinant plasmid was obtained. Conclusion The P. falciparum HRPⅡ gene was subcloned into pEGFP N3 plasmid in the reverse direction and the antisense HRPⅡ / pEGFP N3 fluorescent eukaryotic expression vector was successfully constructed to solve the source of HRPⅡ antisense nucleic acid. In order to further study the anti-malarial effect of antisense HPRⅡ Foundation.