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  • Title: [Molecular cloning and expression of human PDGF-B chain peptide gene and its effect on cat corneal endothelial cell proliferation in vitro].
    Author: Luo WJ, Liu MG, Wang CF, Wang LM.
    Journal: Zhonghua Yan Ke Za Zhi; 2006 May; 42(5):415-9. PubMed ID: 16762235.
    Abstract:
    OBJECTIVE: To construct pET-PDGF-B plasmid with over expression of PDGF and investigate its effects on cat corneal endothelial cells proliferation in vitro. METHODS: Gene rearrangement technique was used to construct the prokaryotic vector with insertion of pET-PDGF-B. The recombinant PDGF-B was obtained from E. coli BL21 (DE3). The effects of recombinant PDGF-B on proliferation and morphologic changes of cultured cat corneal endothelial cells were assayed by modified tertrozalium salt (MTT), inverted phase contrast microscope and transmission electron microscope. RESULTS: PDGF-B peptide gene was inserted into prokaryotic expression vector pET-28a (+), which was confirmed by sequence analysis; a 27,000 protein band was demonstrated as pET-PDGF-B from extraction of E. coli BL21 (DE3) using western blot; MTT assay showed PDGF-BB promoted the proliferation of cat corneal endothelial cells. CONCLUSIONS: A constructed pET-PDGF-B plasmid vector over expression of PDGF is produced. Recombinant PDGF-BB promotes cultured cat corneal endothelial cells proliferation.
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