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目的 构建恶性疟原虫CTP基因的测序重组质粒和真核表达重组质粒 ,以便进一步研究CTP基因的结构与功能。方法 根据已发表恶性疟原虫CTP基因的序列〔1〕,设计并合成一对引物。将扩增的CTP基因的PCR产物连接于测序载体pUC19上 ,经测序反应确定无误。用HindⅢ和BamHⅠ双酶切将CTP基因从测序载体中切下 ,构建于真核表达载体pcDNA3上。 结果 构建了恶性疟原虫CTP基因的测序重组质粒 ,并对其进行了测序。并将恶性疟原虫的CTP基因从测序重组质粒中切下 ,克隆进真核表达重组质粒 pcDNA3。 结论 成功地构建了恶性疟原虫CTP基因的测序重组质粒和真核表达重组质粒 ,为进一步研究其结构与功能奠定了基础
Objective To construct the sequencing recombinant plasmids and eukaryotic recombinant plasmids of Plasmodium falciparum CTP gene in order to further study the structure and function of CTP gene. Methods According to the published sequence of P. falciparum CTP gene [1], a pair of primers was designed and synthesized. The PCR product of the amplified CTP gene was ligated to the sequencing vector pUC19 and the sequencing reaction was confirmed. CTP gene was excised from the sequencing vector by double digestion with Hind III and BamH I, and was constructed on the eukaryotic expression vector pcDNA3. Results A recombinant plasmids of Plasmodium falciparum CTP gene were constructed and sequenced. The CTP gene of Plasmodium falciparum was excised from the sequencing recombinant plasmid and cloned into the eukaryotic expression recombinant plasmid pcDNA3. Conclusion The sequencing recombinant plasmids and eukaryotic recombinant plasmids of Plasmodium falciparum CTP gene were constructed successfully, which laid the foundation for further study of its structure and function