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Cloning and expression of cDNAs from hepatitis E virus structural gene
Bing Ruan1, Edward Zumbika, Shu-Ying Wang
1First Affiliated Hospital, Medical College of Zhejiang University, Hangzhou 310003, China. my_bing@mail.hz.zj.cn
Hepatobiliary & Pancreatic Diseases International : HBPD INT
|November 6, 2003
Summary
Researchers developed a recombinant protein, GST-pORF2, from the hepatitis E virus (HEV) ORF2. This antigen is crucial for creating effective anti-HEV ELISA kits and vaccines against HEV infection.
Area of Science:
- Virology
- Molecular Biology
- Immunology
Background:
- Hepatitis E virus (HEV) infection poses a significant public health challenge.
- Development of diagnostic tools and vaccines against HEV is essential.
Purpose of the Study:
- To produce a recombinant antigen for an anti-HEV ELISA kit.
- To develop a vaccine against hepatitis E virus infection.
Main Methods:
- A 492 bp cDNA fragment from the HEV ORF2 was amplified.
- The fragment was cloned into the pGEX-4T-3 vector and expressed in E. coli.
- Recombinant protein expression was confirmed using Western blot analysis.
Main Results:
- The recombinant plasmid was confirmed by enzyme digestion, PCR, and sequencing.
- A fusion protein (GST-pORF2) of approximately 46 kDa was expressed.
- Western blot analysis showed the fusion protein reacted with anti-HEV positive sera.
Conclusions:
- The recombinant protein GST-pORF2 shows potential for developing anti-HEV diagnostic kits.
- This recombinant antigen may be useful in the creation of a hepatitis E vaccine.