Expression and characterization of the hepatitis E virus ORF3 protein in the methylotrophic yeast, Pichia pastoris

Gene
Sunil K LalShahid Jameel

Abstract

We have used the methylotrophic yeast, Pichia pastoris, to express the open reading frame 3 (ORF3) of the hepatitis E virus (HEV). The ORF3 gene codes for a 123-amino-acid protein that contains highly immunodominant epitopes and is a potentially useful diagnostic and immunoprophylactic antigen. The expressed protein showed positive on immunoblots probed against antibodies raised in rabbit and infected human patient sera. In order to optimize the ORF3 protein expression, we have examined the regulated expression of this protein and characterized it. Unlike its expression in E. coli, the ORF3 protein was present in both the soluble and insoluble fractions of the cell lysate. The expressed protein is not glycosylated and does not undergo any major processing in the host strain.

Citations

Jun 9, 2001·Biochemical and Biophysical Research Communications·S TyagiS K Lal
Aug 4, 2001·Biochemical and Biophysical Research Communications·S TyagiS K Lal
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Dec 13, 2003·Journal of Virology·Milan SurjitSunil K Lal
Jun 28, 2011·Virus Research·Yury Khudyakov, Saleem Kamili
Feb 25, 2011·Virus Research·Imran AhmadShahid Jameel
Jan 1, 2012·Asian Pacific Journal of Cancer Prevention : APJCP·Hossein KeyvaniHamid Reza Mollaie
Aug 17, 2016·EBioMedicine·Hoang van TongThirumalaisamy P Velavan
Jan 2, 2009·Journal of Immunoassay & Immunochemistry·T I UlanovaY E Khudyakov

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