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

Ramakrishnan Nagasundara Ramanan, Wong Bee Tik, Hamid Rajabi Memari, Siti Nor Ani Azaman, Tau Chuan Ling, Beng Ti Tey, Mohd Azmi Mohd Lila, Mohd Puad Abdullah, Raha Abdul Rahim, Arbakariya B. Ariff

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19 Citations (Scopus)

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 pFLAGATS. 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 results suggested 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.

Original languageEnglish
Pages (from-to)285-292
Number of pages8
JournalAfrican Journal of Biotechnology
Volume9
Issue number3
Publication statusPublished - 18 Jan 2010
Externally publishedYes

Keywords

  • Escherichia coli
  • Interferon-α2b
  • mRNA secondary structure
  • Periplasm
  • Signal sequence

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