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Published on: April 3, 2018
[Transient expression of exogenous human platelet-derived growth factor-B in gingival fibroblasts of dog]
Quan Zhong1, Fu-Hua Yan, Yan-Fen Li
1Dept. of Periodontology, The Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou 350002, China.
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 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.
Conclusion:
Eukaryotic expression plasmid carrying hPDGF-B was transfected into gingival fibroblasts successfully, and a kind of fusion protein was expressed.