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

Liu, Y.; Zhao, T.-J.; Yan, Y.-B.; Zhou, H.-M.

Protein Expression and Purification 44(2): 155-161

2005


ISSN/ISBN: 1046-5928
PMID: 15882951
DOI: 10.1016/j.pep.2005.03.030
Accession: 012197931

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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.

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