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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: To study the antigenicity of MBP B165 protein by the expression of the 267 ~ 432 amino acid coding gene fragment of envelope E protein of dengue type 2 strain in China. Methods The gene fragment encoding B165 protein was amplified by PCR and inserted into pMal C2 prokaryotic vector for fusion expression. The expression products were identified by Western blotting and ELISA. The expressed fusion protein MBP B16 5 was used to immunize rabbits and the dengue 2 virus-specific antibody was detected by ELISA. The results showed that the fusion protein MBP B16 5 could specifically bind to Dengue virus type 2 antibody in our country and had high reactivity with the polyclonal antibody of dengue 1,3 and 4 strains. Immunization of rabbits with the expressed protein produced specific antibodies to the E protein of Dengue 2 virus strains in our country, and the antibody cross-reacted with the other three reference strains of virus. 267 through 432 of the amino acid sequence of dengue type 2 virus Conclusion 43 B E protein comprises a certain region including the antigenicity, and reactive epitopes specific subgroup flavivirus, dengue i.e. 4 The structure of the virus conserved epitopes.