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- Title
Transient expression of exogenous human platelet-derived growth factor-B in gingival fibroblasts of dog.
- Authors
ZHONG Quan; YAN Fu-hua; LI Yan-fen; JIANG Yi-ping
- Abstract
Objective To explore transient expression of the eukaryotic expression plasmid carrying human plate-let-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 0PCFM) (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.
- Publication
West China Journal of Stomatology, 2011, Vol 29, Issue 2, p214
- ISSN
1000-1182
- Publication type
Article
- DOI
10.3969/j.issn.1000-1182.2011.02.029