CAJ | 학술논문

目的表达4型戊型肝炎病毒(HEV)开放式阅读框ORF2重组衣壳蛋白并初步建立用于4型HEV早期感染过程研究的细胞模型.方法从猪胆汁中调取4型HEV ORF2的肽段(aa368-606)基因,并且利用大肠杆菌系统得到重组D66蛋白.结果重组D66蛋白在水相中以二聚体为基本单位自组装成分子半径约为13 nm的颗粒.该颗粒与HEV感染的急性期、恢复期患者的血清均有较强的反应性.将D66颗粒与细胞共孵育,发现该颗粒可以与肝实质细胞系HepG2的细胞结合并进入细胞内,这种结合可被恢复期及急性期HEV患者血清所阻断.结论这些结果表明重组D66蛋白颗粒,可以一定程度上模拟4型HEV的表面位点,并且可以特异性吸附并进入细胞,为进一步研究4型HEV感染的分子机制提供基础和平台.
목적 표체4형무형간염병독(HEV)개방식열독광ORF2중조의각단백병초보건립용우4형HEV조기감염과정연구적세포모형.방법 종저담즙중조취4형HEV ORF2적태단(aa368-606)기인,병차이용대장간균계통득도중조D66단백.결과 중조D66단백재수상중이이취체위기본단위자조장성분자반경약위13 nm적과립.해과립여HEV감염적급성기、회복기환자적혈청균유교강적반응성.장D66과립여세포공부육,발현해과립가이여간실질세포계HepG2적세포결합병진입세포내,저충결합가피회복기급급성기HEV환자혈청소조단.결론 저사결과표명중조D66단백과립,가이일정정도상모의4형HEV적표면위점,병차가이특이성흡부병진입세포,위진일보연구4형HEV감염적분자궤제제공기출화평태.
Objective To express the recombinant caspid of genotype 4 hepatitis E virus(HEV) ORF2. Methods HEV recombinant capsid protein D66 was expressed in E. coli, using the ORF2 fragment (aa368-606, obtained from swine bile) of genotype 4 HEV. Results The recombinant capsid proteins D66 self-assemble to be particle with a radius of 13 nm through dimeric form in neutral solution. Coated particles reacted well with sera obtained from patients during acute or recovered phase of HEV infection. Immunofluorescence and immnoblot assay suggested that D66 bound and penetrated HepG2 cell lines, and the process of attachment was blocked by sera collected from patients during acute or recovered phase of HEV infection.Conclusion Recombinant D66 particles simulate the structure at the surface of genotype 4 HEV well and specifically adhere and penetrate the host cells, which lays the foundation for the investigation of the molecular mechanism of genotype 4 HEV infection.