Increase of soluble expression in Escherichia coli cytoplasm by a protein disulfide isomerase gene fusion system

Yang Liu, Tong Jin Zhao, Yong Bin Yan, Hai Meng Zhou

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

Human protein disulfide isomerase (PDI) was selected as a fusion partner to construct a gene expression system to enhance the solubility of recombinant protein in Escherichia coli. DREBIII-1, a plant specific transcriptional factor, was found to mainly form inclusion bodies when expressed in either His-tagged or GST-fusion systems in E. coli. In contrast, when fused with PDI, the expressed DREBIII-1 was in a highly soluble and biologically active form. Two fusion proteins, HDP and HPD, were generated by positioning DREBIII-1 at the N-terminal and C-terminal of PDI, respectively. After purification, HDP exhibited a higher stability and showed only one band on SDS-PAGE, while HPD degraded as several bands. HDP was verified to have the biological function of PDI by isomerase activity assay; meanwhile, it also presented the DNA binding and transcriptional activation characteristic of DREBIII-1 in fluorescence quenching and yeast one-hybrid experiments. The PDI fusion expression system was demonstrated to be highly efficient in generating not only soluble but functional desired proteins.

Original languageEnglish (US)
Pages (from-to)155-161
Number of pages7
JournalProtein Expression and Purification
Volume44
Issue number2
DOIs
StatePublished - Dec 1 2005

Keywords

  • Fusion protein
  • Inclusion bodies
  • Protein disulfide isomerase
  • Soluble expression and purification

ASJC Scopus subject areas

  • Biotechnology

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