我国登革2型病毒43株包膜E蛋白MBP-B165抗原性的研究  

Study on the immunogenicity of the envelope E protein MBP-B165 of Chinese dengue 2 virus 43 strain

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

机构地区：[1]军事医学科学院微生物学流行病学研究所,北京100071

出　　处：《中华微生物学和免疫学杂志》2001年第3期321-323,共3页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金资助项目! (3 93 70 0 3 5 )

摘　　要：目的　通过对我国登革 2型病毒株包膜E蛋白包括B区在内的第 2 6 7～ 432氨基酸编码基因片段的表达 ,研究MBP B16 5蛋白的抗原性。方法　首先采用PCR方法扩增了编码B16 5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。采用蛋白印迹和ELISA法对表达产物进行特异性鉴定。用表达的融合蛋白MBP B16 5免疫大白兔 ,并通过ELISA法检测兔免疫血清中登革2型病毒特异的抗体。结果　表达的融合蛋白MBP B16 5可与我国登革 2型病毒株抗体特异结合 ,而且与登革 1、3和 4型病毒参考株的多克隆抗体均具有较高的反应性。用表达蛋白免疫大白兔可产生针对我国登革 2型病毒株E蛋白的特异抗体 ,并且该抗体与其他 3个型病毒参考株有交叉反应。结论　我国登革 2型病毒 43株E蛋白包括B区在内的第 2 6 7～ 432氨基酸序列具有一定的抗原性 ,而且具有黄病毒亚组特异的反应性表位 ,即 4个型登革病毒的结构保守性表位。Objective The immunogenicity of the envelope E protein MBP-B165 of Chinese D2-43 virus was studied by expressing its gene fragment. Methods The B165 gene fragment was amplified by PCR and then expressed as a fusion protein in E. coli containing the recombinant prokaryotic phagmid pMal-C2 vector. The specificity of the expressed products was proved by the immunoblot (WB) and ELISA. Using the fusion protein MBP-B165 to immunize rabbits, the specific antibodies to dengue 2 virus were detected in the sera of immunized rabbits by ELISA. Results The B165 gene was expressed highly as a fusion protein with the maltose binding protein (MBP) of E. coli. This recombinant protein could react with polyclonal antibodies against dengue virus type 1, 3 and 4. Specific antibodies to dengue 2 virus were detected in the sera of immunized rabbits and were capable of reacting with dengue viruses of other three types. Conclusion The E protein′s 267-432AA of the D2-43 virus, including B domain, possesses the common epitopes of flavivirus subgroup, and this region of E protein from dengue 2 virus contains the structurally conserved epitopes among four types dengue virus.

关 键 词：登革病毒 E基因B区 基因表达 抗原性 

分 类 号：R373[医药卫生—病原生物学]