[Expression of hepatitis E virus structural gene in E. coli].
OBJECTIVE
To obtain recombinant antigen for development of vaccine against hepatitis E virus.


METHODS
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.


RESULTS
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.


CONCLUSIONS
The protein might be useful to develop vaccine against hepatitis E virus infection.
Zhang
M
M
Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052.
Zhao
H
H
Jiang
Y
Y
D000956:Antigens, Viral / Q000096:biosynthesis / Q000235:genetics; D003001:Cloning, Molecular; D000939:Epitopes; D004926:Escherichia coli / Q000235:genetics*; D005814:Genes, Viral / Q000235:genetics*; D016752:Hepatitis E virus / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014761:Viral Hepatitis Vaccines; D015678:Viral Structural Proteins / Q000235:genetics*
D000956:Antigens, Viral; D000939:Epitopes; D011994:Recombinant Proteins; D014761:Viral Hepatitis Vaccines; D015678:Viral Structural Proteins