Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris

Teh, S.H.; Fong, M.Y.; Mohamed, Z. (2011) Expression and analysis of the glycosylation properties of recombinant human erythropoietin expressed in Pichia pastoris. Genetics and Molecular Biology, 34 (3). pp. 464-470.

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    Abstract

    The Pichia pastoris expression system was used to produce recombinant human erythropoietin, a protein synthesized by the adult kidney and responsible for the regulation of red blood cell production. The entire recombinant human erythropoietin (rhEPO) gene was constructed using the Splicing by Overlap Extension by PCR (SOE-PCR) technique, cloned and expressed through the secretory pathway of the Pichia expression system. Recombinant erythropoietin was successfully expressed in P. pastoris. The estimated molecular mass of the expressed protein ranged from 32 kDa to 75 kDa, with the variation in size being attributed to the presence of rhEPO glycosylation analogs. A crude functional analysis of the soluble proteins showed that all of the forms were active in vivo.

    Item Type: Article
    Journal or Publication Title: Genetics and Molecular Biology
    Additional Information: Send correspondence to Zulqarnain Mohamed. Unit of Genetics and Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. E-mail: zulq@um.edu.my.
    Uncontrolled Keywords: erythropoietin, glycosylation, Pichia pastoris, SOE-PCR
    Subjects: R Medicine
    Divisions: Faculty of Medicine
    Depositing User: Fong Mun Yik
    Date Deposited: 11 Jun 2012 09:07
    Last Modified: 11 Jun 2012 09:07
    URI: http://eprints.um.edu.my/id/eprint/3223

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