African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

Effect of promoter strength and signal sequence on the periplasmic expression of human interferon-α2b in Escherichia coli

Ramakrishnan Nagasundara Ramanan1, Wong Bee Tik1, Hamid Rajabi Memari1,2, Siti Nor Ani Azaman1, Tau Chuan Ling3, Beng Ti Tey1,4, Mohd. Azmi Mohd. Lila1, Mohd. Puad Abdullah5, Raha Abdul Rahim1,5 and Arbakariya B. Ariff1,6*
  1Institute of Bioscience, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia. 2Shahid Chamran University, Golestan BLV. Ahvaz, Iran. 3Department of Process and Food Engineering, Faculty of Engineering, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. 4Department of Chemical and Environmental Engineering, Faculty Engineering, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. 5Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. 6Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.  
Email: [email protected]

  •  Accepted: 21 August 2009
  •  Published: 18 January 2010

Abstract

 

Two plasmids, pFLAG-ATS and pET 26b(+), were studied for the periplasmic expression of recombinant human interferon-α2b (IFN-α2b) in Escherichia coli. The pFLAG-ATS contains ompA signal sequence and tac promoter while pET 26b(+) contains pelB signal sequence and T7lac promoter. It was observed that periplasmic expression of IFN-α2b from pET 26b(+) was around 3000 times higher than pFLAG-ATS. Difference in the expression level was attributed to the difference in the promoters and the signal sequences. In silico analysis of mRNA secondary structures were analyzed using Vienna RNA package and MFOLD. The resultssuggested that the increase of expression would mainly due to the difference in the translation initiation associated with secondary structure of mRNA transcribed by both plasmids.

 

Key words: Interferon-α2b, Periplasm, Signal sequence, mRNA secondary structure, Escherichia coli.

Abbreviation

Abbreviation: ompA, outer membrane protein A; pelB, pectate lyase B; PCR, Polymerase chain reaction; ATCC, American type culture collection; RNA, Ribonucleic acid; rRNA, ribosomal RNA; RBS, Ribosomal binding site.

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This article is published under the terms of the Creative Commons Attribution License 4.0

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