恶性疟原虫FCC1/HN株CTP基因真核表达载体的构建(英文)  

CONSTRUCTION OF EUKARYOTIC EXPRESSION VECTOR OF PHOSPHOCHOLINE CYTIDYLYLTRANSFERASE FROM PLASMODIUM FALCIPARUM (FCC1/HN)

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

机构地区：[1]中山医科大学寄生虫学教研室,广东广州510089

出　　处：《中国寄生虫病防治杂志》2002年第1期21-23,共3页Chinese Journal of Parasitic Disease Control

基　　金：The project was supported by" 2 1 1 project"emphasis construction subjectin Guangdong(No.981 69) and fund-ed by doctoral spot of country education committee(No.2 0 0 0 4 5)

摘　　要：目的　构建恶性疟原虫 CTP基因的真核表达载体 ,以便进一步研究其功能。　方法　根据 Genbank已发表恶性疟原虫CTP基因序列 (序列号为 X0 84 0 4 1) ,自行设计并合成了一对引物 ,通过聚合酶链反应扩增出 CTP基因 ,经 Hind 和 Bam H 消化后定向克隆入测序载体 p UC19,构建重组质粒 p UC19- CTP。经双酶切、PCR扩增和序列测定 ,证实插入片段与已知 CTP编码序列完全相同。用 Hind 和 Bam H 消化 p UC19- CTP,将双酶切下的 CTP编码基因片段定向亚克隆入真核表达载体 pc DNA3。　结果经双酶切和 PCR扩增鉴定证实 CTP基因正向插入真核表达载体 pc DNA3中。　结论　真核表达质粒 pc DNA3-Objective To construct the eukaryotic expression plasmid of phosphocholine cytidylyltransferase (CTP) gene of Plasmodium falciparum in order to study on its function. Methods According to the gene sequence of phosphocholine cytidylytransferase of P. falciparum (Genbank accession No. X84041), a pair of primers were designed and synthesized. The coding sequence of CTP was amplified and cloned into pUC19 plasmid. After sequencing, the gene was subcloned into eukaryotic expression vector pcDNA3. Recombinant plasmid was analysed by restriction enzyme digestion and polymerase chain reaction. Results CTP gene was correctly cloned into eukaryotic expression vector pcDNA3 by identification of restriction enzyme digestion and polymerase chain reaction. Conclusion The eukaryotic expression plasmid pcDNA3/CTP was successfully constructed.

关 键 词：恶性疟原虫 聚合酶链反应 真核表达载体 基因表达 构建 

分 类 号：R382.31[医药卫生—医学寄生虫学]