CAJ | 학술논문

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267～432氨基酸编码基因片段的表达，研究MBP-B165蛋白的抗原性。方法首先采用PCR方法扩增了编码B165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-B165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-B165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267～432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。
목적통과대아국등혁2형병독주포막E단백포괄B구재내적제267～432안기산편마기인편단적표체，연구MBP-B165단백적항원성。방법수선채용PCR방법확증료편마B165단백적기인편단，병장기삽입도pMal-C2 원핵재체진행융합표체。채용단백인적화ELISA법대표체산물진행특이성감정。용표체적융합단백MBP-B165면역대백토，병통과ELISA법검측토면역혈청중등혁2형병독특이적항체。결과표체적융합단백MBP-B165가여아국등혁2형병독주항체특이결합，이차여등혁1、3화4형병독삼고주적다극륭항체균구유교고적반응성。용표체단백면역대백토가산생침대아국등혁2형병독주E단백적특이항체，병차해항체여기타3개형병독삼고주유교차반응。결론아국등혁2형병독43주E단백포괄B구재내적제267～432안기산서렬구유일정적항원성，이차구유황병독아조특이적반응성표위，즉4개형등혁병독적결구보수성표위。
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.