我国登革2型病毒株E蛋白B抗原区基因的表达与鉴定  

Expression and identification of the E gene B domain of dengue2 virus isolated in China

作　　者：仝莉莉[1] 秦鄂德[1] 李同据[1] 杨佩英[1] 于曼[1] 

机构地区：[1]军事医学科学院微生物学流行病学研究所

出　　处：《军事医学科学院院刊》1999年第2期90-93,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家自然科学基金

摘　　要：目的和方法：通过对登革２型病毒Ｅ蛋白Ｂ区基因片段的表达，研究Ｂ区蛋白的抗原性。首先采用ＰＣＲ方法扩增了编码Ｂ区蛋白的基因片段，并将其插入到ｐＭａｌ－Ｃ２原核载体进行融合表达。采用蛋白质印迹法和ＥＬＩＳＡ法对表达产物进行特异性鉴定。结果与结论：在构建的重组质粒Ｂ１６５－ｐＭａｌ中，Ｅ蛋白Ｂ区与大肠杆菌的麦芽糖结合蛋白（ｍａｌｔｏｓｅｂｉｎｄｉｎｇｐｒｏｔｅｉｎ，ＭＢＰ）基因以融合形式高效表达。该融合蛋白可与登革１～４型病毒的鼠腹水抗体起特异反应，表明我国登革２型病毒株Ｅ蛋白Ｂ区具有黄病毒亚组反应性表位。Objective and Methods: To study the antigenicity of the Eglycoprotein B domain of dengue 2 virus by expressing its gene fragment. The B domain gene fragment was amplified by means of PCR, then expressed as a fusion protein in TBX]E.coli containing the recombinant prokaryotic phagemid pMalC2 vector. The specificity of the expressed products was identified by the immunoblot (WB) and ELISA. Results and Conclusion: The B domain gene was expressed highly as a fusion protein with the maltose binding protein of E.coli . This recombinant protein reacted with hyperimmune mouse ascitic fluid against the four serotypes of dengue virus. These results suggest that common antigenic epitopes to the flavivirus subgroup be present on the Eglycoprotein B domain of the dengue2 virus isolated in China.

关 键 词：登革热病毒 E基因B区 基因表达 ELISA 

分 类 号：R373.33[医药卫生—病原生物学]