Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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A Novel Expression System for Recombinant Marine Mussel Adhesive Protein Mefp1 Using a Truncated OmpA Signal Peptide

  • Lee, Sang Jun (Biotechnology Research Institute, National Fisheries Research and Development Institute) ;
  • Han, Yun Hee (Biotechnology Research Institute, National Fisheries Research and Development Institute) ;
  • Nam, Bo Hye (Biotechnology Research Institute, National Fisheries Research and Development Institute) ;
  • Kim, Young Ok (Biotechnology Research Institute, National Fisheries Research and Development Institute) ;
  • Reeves, Peter R. (Department of Microbiology, University of Sydney)
  • Received : 2008.03.04
  • Accepted : 2008.04.29
  • Published : 2008.07.31
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Abstract

To express an increased level of recombinant Mefp1 (marine mussel adhesive protein) in soluble form, we constructed expression vectors encoding truncated OmpA signal peptide-Mefp1 fusion proteins. OmpA signal peptide (OmpASP) is the 21 residue peptide fragment of the 23 residue OmpA signal sequence cleavable by signal peptidase I. We successfully produced increased levels of soluble recombinant Mefp1 (rMefp1) with various deletions of OmpASP, and found that the increased expression was caused by the increased pI of the N-terminus of the fusion proteins (${\geq}10.55$). All the OmpA signal peptide segments of 3-21 amino acids in length had the same pI value (10.55). Our results suggest that the pI value of the truncated OmpASP ($OmpASP_{tr}$) play an important role in directional signaling for the fusion protein, but we found no evidence for the presence of a secretion enhancer in OmpASP. For practical applications, we increased the expression of soluble rMefp1 with $OmpASP_{tr}$ peptides as directional signals, and obtained rMefp1 with the native amino terminus (nN-rMefp1) using an $OmpASP_{tr}$ Xa leader sequence that contains the recognition site for Xa protease.

Keywords

Acknowledgement

Supported by : National Fisheries Research and Development Institute

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