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| Cloning and expression of cDNAs from hepatitis E virus structural gene |
| Bing Ruan, Edward Zumbika, Shu-Ying Wang, Yong Chen, Yi-Lin Ma, Ya-Gang Chen and Ke-Zhou Liu |
Hangzhou, China
From the First Affiliated Hospital, Medical College of Zhejiang University, Hangzhou 310003, China (Ruan B, Zumbika E, Ma YL, Chen YG and Liu KZ); Hangzhou First People’s Hospital (Wang SY); Zhejiang Academy of Medical Sciences (Chen Y)
Correspondence: Bing Ruan, MD, First Affiliated Hospital, Medical College of Zhejiang University, Hangzhou 310003, China (Tel: 86-571-87236755; Fax: 86-571-87236755; Email: my_bing@mail.hz.zj.cn) |
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Abstract OBJECTIVE: To obtain recombinant antigen for development of anti-HEV ELISA kit and vaccine against hepatitis E virus infection.
METHODS: A 492 base cDNA was collected from 5’-terminus of open reading frame 2 (ORF2) among epidemic hepatitis E virus (HEV) isolated from Xinjiang, Western China. The fragment was digested with BamH Ⅰ and EcoRⅠ, and inserted into vector pGEX-4T-3 which was also digested by the same enzyme. The recombinant plasmid was transformed into E.coli TG-1 and the fusion protein expressed was confirmed by Western blot analysis using serum of a patient with hepatitis E.
RESULTS: The recombinant plasmid was identified and confirmed with enzyme digestion, polymerase chain reaction (PCR) and sequencing, respectively. A protein band of about 46 kDa was demonstrated by SDS-PAGE and designated GST-pORF2. The result of Western blot analysis suggested that the fusion protein reacted with anti-HEV positive sera at a dilution of 1∶100.
CONCLUSION: The recombinant protein GST-pORF2 may be useful in developing anti-HEV ELISA kit and vaccine against hepatitis E virus infection.
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2003, Vol. 2 
