CJCU

Chem. J. Chinese Universities ›› 2012, Vol. 33 ›› Issue (11): 2481.doi: 10.7503/cjcu20120368

• Biological Chemistry • Previous Articles     Next Articles

Expression and Characterization of a Recombinant Truncated Capsid Protein of Hepatitis E Virus

LI Xiao1,2, QU Yong-Gang2,3, JIA Peng2,4, LIU Li-Ming2,5, JI Hui-Fan1,2, HE Dong-Yun1,2, JIN Ning-Yi2, CHI Bao-Rong1   

  1. 1. The First Hospital of Bethune Faculty of Medical Sciences, Jilin University, Changchun 130021, China;
    2. Laboratory of Genetic Engineering of PLA, Academy of Military Medical Sciences, Changchun 130122, China;
    3. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China;
    4. Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen 518000, China;
    5. College of Veterinary Medicine, Jilin Academy of Agricultural Science and Technology, Jilin 132101, China
  • Received:2012-04-17 Online:2012-11-10 Published:2012-10-15

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Abstract:

Hepatitis E is an enterically transmitted viral disease caused by infection with hepatitis E virus(HEV). HEV is a nonenveloped virus classified in the family of Caliciviridae. The virus appears to be a polyadenylated, positive-stranded RNA virus with three major open reading frames(ORFs). The capsid protein of HEV is encoded by the open reading frame 2(ORF2). We attempted to produce a truncated capsid protein, designed p293, in E. coli. The p293 gene encoding amino acids aa382-aa674 of HEV ORF2 was designed based on the full length of HEV ORF2, cloned into the vector pET28a, and expressed in E. coli strain BL21(DE3). SDS-PAGE and Western blot analysis demonstrated that the recombinant protein p293 could well expressed in E. coli. Under optimized conditions(1 mmol/L IPTG, 250 r/min, 37℃, 5 h), the yield of soluble p293 was approximately 66.15% of the total protein. It was observed, by transmission electron microscopy, that p293 expressed in E. coli formed 30-40 nm viral-like particles. The experimental results also showed that p293 reacted with HEV positive serum and had a good reactionogenicity. These results showed that the p293 may has utility in the analysis of cell specific factors in the protein processing and assembly of HEV, and serve as useful antigen for both diagnostic and vaccine purposes.

Key words: Hepatitis E virus(HEV), Capsid Protein, Viral-like particle(VLP), Prokaryotic expression

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