Research Article Open Access

Molecular Cloning and Heterologous Expression of Human Interferon Alpha2b Gene

I. Made Artika1, Yemima Budirahardja2 and Anna Lucia Ekowati3
  • 1 Eijkman Institute for Molecular Biology, Indonesia
  • 2 National University of Singapore, Singapore
  • 3 Atma Jaya Catholic University of Indonesia, Indonesia

Abstract

Human alpha Interferons (hIFNα) have been shown to have antiviral, antiproliferative and immunomodulatory activities. The human interferon alpha2b (hIFNα2b), is one of the human interferon alpha2 sub variants, naturally synthesized as a polypeptide of 188 amino acid residues, the first 23 residues of which represents a signal peptide. In the present study, the hIFNα2b gene was expressed after being fused with Glutathione S-Transferase (GST) gene. The hIFNα2b gene was amplified from human genomic DNA by using a pair of specific primers, cloned into an Escherichia coli expression vector and expressed in E. coli cells under the direction of the tac promoter. The expressed protein was purified using a one-step affinity chromatography column containing immobilized gluthatione-bound resin. The purified protein was shown to react specifically with anti-human-interferon-alpha antibody, confirming that the protein was the human interferon alpha molecule. This strategy has the potential to be used as an alternative mean for production of pure human interferon α proteins for therapeutic purposes and for further studies on their molecular characterization and mechanism of action.

American Journal of Biochemistry and Biotechnology
Volume 9 No. 4, 2013, 423-429

DOI: https://doi.org/10.3844/ajbbsp.2013.423.429

Submitted On: 13 August 2013 Published On: 20 December 2013

How to Cite: Artika, I. M., Budirahardja, Y. & Ekowati, A. L. (2013). Molecular Cloning and Heterologous Expression of Human Interferon Alpha2b Gene. American Journal of Biochemistry and Biotechnology, 9(4), 423-429. https://doi.org/10.3844/ajbbsp.2013.423.429

  • 3,694 Views
  • 4,076 Downloads
  • 6 Citations

Download

Keywords

  • Recombinant Human Interferon Alpha2b
  • Escherichia coli
  • Protein Expression
  • Protein Purification