Purification and Expression of ORF2 Protein of Hepatitis E Virus in E.coli

Hepatitis E virus (HEV) is a causative agent of acute hepatitis among people of different age groups and has high mortality rate of up to 30% among pregnant women. Therefore, primary prevention of HEV infection is essential. The aim of this study was to obtain the highly purified (ORF2) protein, which might be a future HEV vaccine candidate. The truncated orf2 gene (orf2.1), encoding the 112-660 amino acid of HEV capsid protein sequence, was optimized, synthesized. After induction with isopropyl-β-D-thiogalactopyranoside (IPTG) and optimizing the conditions of expression, The target protein was highly expressed and purified by Ni(2+)-che-late affinity chromatography. The purified protein was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.The results obtained from optimizing the expression con-ditions showed℃that the highest expression of the protein was obtained by adding IPTG at a final concentration of 1 mM at 37 for four hours. The expression and purification of truncated ORF2 protein was confirmed by SDS-PAGE and western blotting. SDS-PAGE analysis showed a protein band of about 55 kDa. SDS-PAGE of the purified protein revealed that the highest amount of target protein in elution buffer at the pH of 4.5 was obtained. The yield of the purified protein was about 1 mg/L of culture media

کلیدواژه ها:

Purification ، Expression ، ORF2 Protein ، Hepatitis E virus ، E.coli