关键词: 血小板源性生长因子-B, 基因转染, 牙龈成纤维细胞

Abstract:

Objective To explore transient expression of the eukaryotic expression plasmid carrying human platelet- derived growth factor-B（hPDGF-B） in gingival fibroblasts of Beagle dog. Methods Plasmid carrying hPDGF-B （EX-A0380-M03） was amplified and identified, and then transfected into gingival fibroblasts of Beagle dog. Reverse transcription polymerase chain reaction（RT-PCR）, immunocytochemistry, enzyme-linked immunosorbent assay（ELISA） and Western bolt were choose to detect the expression of hPDGF-B. Results Target gene carried by EX-A0380- M03 was hPDGF-B. Green fluorescene protein（GFP） expressed by transfected gingival fibroblasts was observed under inverted phase contrast fluorescence microscope（IPCFM）（after 24 hours） and the transfection efficiency was 18%-38% （after 48 hours）. Serials other methods（RT-PCR, immunocytochemistry, and ELISA） mentioned above also convinced that cells expressed hPDGF-B, and the protein that was a kind of fusion protein composed of PDGF-BB and GFP was idenified by Western blot. Conclusion Eukaryotic expression plasmid carrying hPDGF-B was transfected into gingival fibroblasts successfully, and a kind of fusion protein was expressed.

Key words: platelet-derived growth factor-B, gene transfection, gingival fibroblasts