Production of Human Alpha 2b Interferon in E.coli Expression System

Document Type : Original Article

Authors

1 Department of Agricultural Biotechnology, Payam Nur University, Karaj, Iran.

2 Department of Agronomy and Plant Breeding, Shahid Chamran University, Ahvaz, Iran

3 Department of Biology, Shahid Chamran University, Ahvaz, Iran

4 Department of Agricultural Biotechnology, Payam Nur University,Tehran, Iran.

Abstract

Background and Objective: Interferons are part of the body's defense system against viruses. These proteins are generated in the body, and then they trigger the cells around them to produce inhibitor proteins against virus replication. The purpose of this study was cloning and expression of the human interferon alpha-2b gene in preplasmic space of Escherichia coli.
Subjects and Methods: In this research, Alpha 2b interferon gene was cloned in a pET-26b (+) expression vector, under the control of T7 promoter and pelB periplasmic signal peptide sequence using NcoI and XhoI restriction enzymes. The expression vector was transformed into Escherichia colistrain BL21 (DE3).Cloning of alpha interferon gene confirmed using colony PCR, digestion and DNA sequencing.Gene expression of alpha interferon was examined by SDS-PAGE and Dot blot analysis. Also the possibility of periplasmic targeting was determined using SDS-PAGE and dot blot analysis at 15 hrs after induction.
Results: The results showed that Alpha 2b interferon was produced in bacterial expression system and targeted to periplasmic space.
Conclusion: Our study showed that the production of pharmaceutical recombinant protein in preplasmic space of bacterial expression system is fissible and can be ustilized as a cheaper source and more efficient than conventional expression systems.

Keywords

References

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Jundishapur Scientific Medical Journal
Volume 12, Issue 3 - Serial Number 84
July and August 2013
Pages 325-334

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History

  • Receive Date: 10 July 2011
  • Revise Date: 13 February 2013
  • Accept Date: 19 February 2013

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