丙型肝炎病毒抗原决定簇与霍乱毒素B亚基的融合表达以及融合蛋白的抗原性  

EXPRESSION AND ANTIGENICITY OF CHIMERIC PROTEINS OF CHOLERA TOXIN B SUBUNIT WITH HEPATTTIS CVIRUS EPITOPES Cal

作　　者：曹诚[1] 马清钧[1] 石成华[1] 于公义[1] 李平[1] 李杰之[1] 张艳红[1] 张京生[1] 

机构地区：[1]北京军事医学科学院生物工程研究所

出　　处：《病毒学报》1995年第2期99-106,共8页Chinese Journal of Virology

摘　　要：通过固相化学合成法，合成了编码丙型肝炎病毒（ＨＣＶ）结构区和非结构区的４个抗原决定簇基因，这些抗原决定簇基因片段以不同方式串朕后与ｃｔｘＢ基因融合，构建了１２种表达不同嵌合蛋白的重组质粒。各重组质粒转化大肠杆菌后均能高效分泌性表达融合蛋白，表达产量随所融合的抗原决定簇不同在１０～５０μｇ／ｍｌ之间，表达水平主要与抗原决定簇的氨基酸组成有关，而与抗原决定簇的大小及串联次数关系不大。融合蛋白通过亲和层析纯化达到了电泳纯。对１１种融合蛋白中ＨＣＶ抗原决定簇的反应原性的研究表明，多数融合蛋白中ＨＣＶ抗原决定簇均能与对应的ＨＣＶ抗体结合。其中以融合蛋白９５０８２为抗原研制的抗－ＨＣＶＥＬＩＳＡ试剂具有良好的灵敏度和特异性。Four oligonucleotides,which encode epitopes of structural and nonstructural antigens of HCV,were chemically synthesized and fused with cholera toxin B subunit(CTB)gene in diferentways.Twelve recombinant plasmids were constructed and all of them could express fusion proteins consisting of CTB and HCV epitopes.The expression level of fusion proteins ranged from 10-50μg/ml depending on amino acid composition of the epitopes and more than 95%of thechimeras were secreted into medium.Purified fusion proteins were obtained by affinity chromatography.The reactivity of the 11 chimeras with human anti-HCV positive sera was evaluated in this work.The results demonstrated that most of the chimeras could react with anti-HCV antibodies in the sera.TheELISA kit showed high accordance with ABBOTT An-ti-HCV ELISA kit by using fusion protein 95082 as antigen.

关 键 词：丙型肝炎病毒 霍乱毒素B亚基 基因融合 

分 类 号：R373.21[医药卫生—病原生物学]