利用RT-PCR的方法检测恶性疟原虫海南株FCC1/HN CT基因在红细胞内期的表达及CTP基因真核表达载体的构建(英文)  

Detection of CTP gene expression in the asexual erythrocytic stages of Plasmodium falciparum (FCC1/HN) by reverse Transcriptase-polymerase chain reaction and construction of eukaryotic expression vector of CTP gene

作　　者：陈慧红[1] 余新炳[1] 吴忠道[1] 徐劲[1] 陆家海[1] 

机构地区：[1]中山医科大学寄生虫教研室,广州510089

出　　处：《热带医学杂志》2001年第1期10-12,共3页Journal of Tropical Medicine

摘　　要：目的 利用反转录ＰＣＲ（ＲＴ－ＰＣＲ）的方法检测ＣＴＰ基因是否在恶性疟原虫红内期表达，并构建ＣＴＰ因的真核表达载体，以便进一步研究其功能。方法 按常规方法体外培养红内期恶性疟原虫，用Ｔｒｉｚｏｌ试剂提取红内期疟原虫总ＲＮＡ．通过ＲＴ－ＰＣＲ方法扩增恶性疟原虫ＦＣＣ１／ＨＮ珠ＣＴＰ Ｍ码基因并构建ＣＴＰ基因的真核表达载体。结果 获得了恶性疟原虫ＣＴＰｃＤＮＡ的全编码区序列并将其克隆入真核表达载体ｐｃＤＮＡ３。结论（ＣＴＰ基因在恶性疟原虫ＦＣＣ１／ＨＮ株红细胞内期表达并成功地构建了ＣＴＰ基因的重组表达质粒。Objective To detect whether the CTP(phosphocholine cytidylyltransferase) gene was expressed in the asexual erythrocytic stages of Plasmodium falciparum (FCC1 / HN ) by using the RT- PCR and to construct eukaryotic expression vector of CTP. Method The erythrocytic stage parasites of Plasmodium falciparum were cultured as described by Trager and Jensen. RNA from erythrocytic stage parasite was extracted by using Trizol reagent. The complete genes coding for CTP gene isolates FCCI /HN were amplified by reverse transcriptase- polymerase chain reaction(RT-PCR). CTP gene was cloned into eukaryotic expression vector pcDNA3. Results CTP encoding gene was amplified from the erythrocytic stages of Plasmodium falciparum (FCC1 / HN) and eukaryotic expression vector of CTP was constructed. Conclusion CTP gene was expressed in the erythrocytic stages of Plasmodium falciparum(FCCI /HN) and eukaryotic expression vector of CTP was successfully constructed.

关 键 词：恶性疟原虫 反转录聚合酶链式反应 磷酸胆碱胞苷酰转移酶 CTP 基因 基因真核表达载体 红细胞 

分 类 号：R382.31[医药卫生—医学寄生虫学]