Transient expression of exogenous human platelet-derived growth factor-B in gingival fibroblasts of dog
Zhong, Q.; Yan, F.-H.; Li, Y.-F.; Jiang, Y.-P.
Hua Xi Kou Qiang Yi Xue Za Zhi 29(2): 214-219
2011
ISSN/ISBN: 1000-1182 PMID: 21598503 Document Number: 651306
To explore transient expression of the eukaryotic expression plasmid carrying human platelet-derived growth factor-B (hPDGF-B) in gingival fibroblasts of Beagle dog. 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. Target gene carried by EX-A0380-M03 was hPDGF-B. Green fluorescence 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 identified by Western blot. Eukaryotic expression plasmid carrying hPDGF-B was transfected into gingival fibroblasts successfully, and a kind of fusion protein was expressed.