弓形虫P30基因原核表达载体的构建  

CONSTRUCTION OF THE PROKARYOTIC EXPRESSION VECTOR OF THE MAJOR SURFACE ANTIGEN P30 GENE OF TOXOPLASMA GONDII

作　　者：丛华[1] 古钦民[1] 赵跃然[2] 游力[2] 黄勇[1] 王伟[1] 

机构地区：[1]山东医科大学寄生虫学教研室,山东济南250012 [2]山东省医学科学院基础医学研究所

出　　处：《中国寄生虫病防治杂志》2000年第3期172-174,共3页Chinese Journal of Parasitic Disease Control

基　　金：山东省卫生厅资助项目

摘　　要：为获取具有生物学活性的弓形虫 P30蛋白 ,采用定向克隆的方法 ,自行设计引物通过 PCR扩增得到 P30基因片段 ,用 Eco R 和 Sal 双酶切后 ,连接到同样双酶切的原核表达载体 (p BV2 2 0和 p MAL P2 )上 ,转化大肠杆菌 DH5 α,分别得到含重组质粒 p BV2 2 0 - P30和 p MAL P2 - P30的工程菌。扩菌提取质粒经过酶切分析和 PCR扩增鉴定后 ,证实 P30基因的两个原核表达载体构建成功 。To obtain biological activity of P30 protein of Toxoplasma gondii , we constructed two prokaryotic expression vector contained a gene encoding P30 gene T. gondii . A pair of primes were designed and synthesized according to the published gene sequence of the P30 gene. At the 5′ end of sens and antisens strand of primers EcoRⅠ and SalⅠ enzyme sites were added respectively. Using PCR, the P30 gene which has 976 bp was amplified from genomic of RH strain T. gondii . After digested by EcoRⅠ and SalⅠ, the amplified gene fragments and plasmid vector(one is non fusion type vector pBV220,the other is fusion type vector pMALP 2) were ligated to construct recombinant plasmids. Then the ligation were transferred into E. coli DH5α. The positive clones were primarily confirmed by anti ampicillin selecting, restriction enzymes digestion and PCR. The successful construction of prokaryotic expression vector would be a good basis for the expression of P30 protein.

关 键 词：弓形体 P30基因 克隆 

分 类 号：R382.5[医药卫生—医学寄生虫学]