间日疟原虫乳酸脱氢酶GST融合表达载体的构建及表达  

Cloning and expression of lactate dehydrogenase of Plasmodium vivax

作　　者：周志成[1] 孙莉[1] 吴英松[1] 廖小青[1] 郝文波[1] 

机构地区：[1]南方医科大学生物技术学院抗体工程研究所,广东广州510515

出　　处：《分子诊断与治疗杂志》2010年第2期91-93,共3页Journal of Molecular Diagnostics and Therapy

基　　金：国家科技重大专项(2008ZX10004-007);广东省医学科学技术研究基金项目(A2006365);广州市科技计划项目(2002Z3-E4012)

摘　　要：目的构建间日疟原虫乳酸脱氢酶(PvLDH)融合表达载体,并表达PvLDH的GST融合蛋白。方法将PvLDH基因片段克隆到表达载体pGEX-4T-1中,构建PvLDH/pGEX-4T-1融合表达载体,IPTG诱导表达目的基因,SDS-PAGE电泳分析表达产物,Western blot检测其抗原性。结果成功构建了PvLDH/pGEX-4T-1融合表达系统,在大肠杆菌BL21中以包涵体形式高效表达,表达产物能与恶性疟原虫和间日疟原虫感染患者血清反应,而不与正常人血清反应。结论间日疟原虫LDH蛋白在大肠杆菌中获得高效表达,表达产物具有良好的抗原性。Objective To clone and express lactate dehydrogenase （LDH） of Plasmodium vivax. Methods PvLDH gene was cloned into pGEX-4T-1 vector. The recombinant plasmid was transformed and then induced to express in the E. coli BL21. The expressed product was analyzed by SDS-PAGE and Western blot. Results The recombinant protein GST-PvLDH was expressed as an insoluble fusion protein with GST,and it exhibited a specific reaction with immune sera obtained from patients infected by Plasmodium falciparum or Plasmodium vivax,but not with sera from healthy persons. Conclusion PvLDH has been successfully expressed as GST fusion protein and the expressed protein has certain antigencity.

关 键 词：间日疟原虫 乳酸脱氢酶 表达 

分 类 号：R382.31[医药卫生—医学寄生虫学]