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

32 Citations (Scopus)

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 2005

Fingerprint

Protein Disulfide-Isomerases
Gene Fusion
Escherichia coli
Cytoplasm
Fusion reactions
Genes
Isomerases
Inclusion Bodies
Recombinant Proteins
Gene expression
Yeast
Solubility
Transcriptional Activation
Purification
Polyacrylamide Gel Electrophoresis
Quenching
Assays
Proteins
Yeasts
Fluorescence

Keywords

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

ASJC Scopus subject areas

  • Biochemistry

Cite this

Increase of soluble expression in Escherichia coli cytoplasm by a protein disulfide isomerase gene fusion system. / Liu, Yang; Zhao, Tong Jin; Yan, Yong Bin; Zhou, Hai Meng.

In: Protein Expression and Purification, Vol. 44, No. 2, 12.2005, p. 155-161.

Research output: Contribution to journalArticle

Liu, Yang ; Zhao, Tong Jin ; Yan, Yong Bin ; Zhou, Hai Meng. / Increase of soluble expression in Escherichia coli cytoplasm by a protein disulfide isomerase gene fusion system. In: Protein Expression and Purification. 2005 ; Vol. 44, No. 2. pp. 155-161.
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