期刊
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
卷 47, 期 -, 页码 105-112出版社
WILEY
DOI: 10.1042/BA20060114
关键词
burn-wound repairing; epitheliotropic effects; hexahistidine fusion protein; human connective tissue growth factor (hCTGF); human umbilical-vein endothelial cell ctgf gene cloning; rhesus monkey (Maccca mulatta)
CTGF (connective-tissue growth factor) has been characterized as an extracellular-matrix-associated protein that modulates basic-fibroblast-growth-factor signalling and angiogenesis. In the present paper, the cloning of the ctgf gene from human umbilical-vein endothelial cells and expression of the protein in Escherichia coli as an N-terminal hexahistidine fusion protein is described. Recombinant human CTGF (rhCTGF) was expressed and purified so that we could investigate its effect on the proliferation of human embryo fibroblast KMB-17 and NIH3T3 cells. The results indicated not only that the protein was properly folded, but also that it had the same specific activity and stability as the native protein. Furthermore, we administered this recombinant protein in a non-human primate [rhesus monkey (Macaca mulatta)] burn-wound model and report the clinical findings and structural effects. Epitheliotrophic effects were conspicuous in wounded tissues at 10-100 ng of CTGF/cm(2), suggesting that administered rhCTGF can play a normal physiological role in wound repairing in a nonhuman primate model.
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