恶性疟原虫海南株CTP基因测序重组质粒及真核表达重组质粒的构建

Construction of sequencing recombinant plasmid and eukaryotic expressionrecombinant plasmid of CTP(phosphocholine cytidylytransferase)from Plasmodium falciparum(FCC1/HN)

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 。

Aim To construct the eukaryotic expression vector of CTP gene,and study on its function Methods According to the published CTP gene sequence,design and synthesize a pair of pimers CTP gene was amplified by PCR and cloned into sequencing vector pUC19 Sequence of CTP gene is proved by sequencing reaction After CTP gene was digested with HindIII and BamHI,CTP gene was ligated to eukaryotic expression vector pcDNA3 Results Sequencing vector of CTP gene was successfully constructed Sequence of CTP gene was proved correct by sequencing reaction CTP gene was successfully inserted in the eukaryotic expression vector pcDNA3 from sequencing vector pUC19 CTP Conclusion Recombinant sequencing plasmid pUC19 CTP and recombinant expression plasmid pcDNA3 CTP can be successfully constructed