Expression of hepatitis e virus structural gene in E. coli
Zhang, M.; Zhao, H.; Jiang, Y.
Zhonghua Shi Yan he Lin Chuang Bing du Xue Za Zhi 13(2): 130-132
1999
ISSN/ISBN: 1003-9279 PMID: 12569778 Document Number: 508880
To obtain recombinant antigen for development of vaccine against hepatitis E virus. Amplified the structural gene (5,816-7,126 nt) by PCR. The upstream primer was 5'-CCATATGAATTCAATAACCTC-3' and the downstream primer was 5'-GGGATCCTATAACTCCCGAGT-3'. Cut the PCR product with Nde I and BamHI, then inserted this fragment into the plasmid pET-11 where a cut was by the same restriction endonucleases. The expression plasmid named pEa47 was transformed into E. Coli BL21. The recombinant strains were grown at 37 degrees C and induced by IPTG. The recombinant protein was confirmed by Western blot analysis using serum from hepatitis E patient. The structural gene of hepatitis E virus from open reading frame 2,224-660 aa, was expressed in E. Coli BL21. Western blot assay showed that the expressed 50,000 recombinant protein specifically reacted with the serum antibody from the hepatitis E patient. The protein might be useful to develop vaccine against hepatitis E virus infection.