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

被引：36
作者：
Liu, Y
Zhao, TJ
Yan, YB [1 ]
Zhou, HM
机构：
[1] Tsing Hua Univ, Dept Biol Sci & Biotechnol, Beijing 100084, Peoples R China
[2] Tsing Hua Univ, State Key Lab Biomembrane & Membrane Biotechnol, Beijing 100084, Peoples R China
来源：
PROTEIN EXPRESSION AND PURIFICATION | 2005年 / 44卷 / 02期
关键词：
protein disulfide isomerase; inclusion bodies; soluble expression and purification; fusion protein;
D O I：
10.1016/j.pep.2005.03.030
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Human protein disulfide isornerase (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. (c) 2005 Elsevier Inc. All rights reserved.
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收藏
页码：155 / 161
页数：7
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