日本血吸虫中国大陆株28kDa GST基因在大肠杆菌中的表达  

EXPRESSION IN E. COLI OF 28kDa GLUTATHIONE S-TRANSFERASE GENE FROM CHINESE STRAIN OF SCHISTOSOMA JAPONICUM

作　　者：田锷[1] 杨冠珍[2] 蔡幼民 沈纬[1] 施福恢[1] 吴祥甫[2] 

机构地区：[1]中国农业科学院上海家畜寄生虫病研究所,上海200232 [2]中国科学院上海生物化学研究所,上海200031

出　　处：《动物学报》1996年第4期421-427,共7页ACTA ZOOLOGICA SINICA

基　　金：总理专项基金(94-Y-40)资助的课题

摘　　要：在大肠杆菌ＴＢ１中表达合日本血吸虫中国大陆株２８ｋＤｅ抗原基因的重组质粒，表达产物是融 合蛋白，分子量为３３ｋＤａ。采用谷胱甘肽琼脂糖亲和层析柱纯化表达产物。２、４－二硝基氯苯／谷胱 甘肽分光光度测定法和琼脂糖－淀粉凝胶电泳显示重组抗原具有较高的谷胱甘肽Ｓ－转移酶（ＧＳＴ） 活力。酶联免疫吸附试验（ＥＬＩＳＡ）结果表明该重组抗原可检测到重组２８ｋＤａ ＧＳＴ或日本血吸虫 虫体ＧＳＴ免疫绵羊血清中的特异性抗体；免疫印迹试验（Ｗｅｓｔｅｒｎ Ｂｌｏｔｔｉｎｇ）分析揭示重组抗原的 ３３ｋＤａ抗原分子能被这两种免疫血清识别，证明它具有ＧＳＴ抗原性。This paper reports the expression of a recombinant plasmid which contains the 28kDa glutathione S-transferase (GST) gene of the Chinese strain of Schistosoma japonicum in Es- cherichia coli TB1. The product was a 33kDa fusion protein, which was purified by glu- tathione-agarose affinity chromatography. The purified recombinant antigen showed GST activity detected by 1-chloro-2, 4-dintrobenzene /glutathione spectrophotometry and a- garose-starch gel electrophoresis. The result of ELISA showed that the recombinant antigen could detect specific antibodies of sheep serum immunized with recombinant 28kDa GST or with native GST from the Chinese strain of Schistosoma japonicum, and the 33kDa molecule of recombinant antigen could be recognized by Western Blotting. All these results provide evidence that the recombinant antigen had GST antigenicity, and it would be assessed as a candidate vaccine in animal experiment against Schistosomiasis japonica.

关 键 词：日本血吸虫 中国大陆株 谷胱甘肽S 转移酶 基因 

分 类 号：R383.24[医药卫生—医学寄生虫学]