Objective: To construct the recombined retroviral expression vector of BMP2/pLEGFP and investigate the bio-activity of the expressed chimeric protein.
Methods: The recombinant vector constructed by gene recombinant technology was analyzed by restriction enzyme digestion and PCR. BMP2/pLEGFP was transfected into COS-7 cells with liposome transfection reagents for transient expression. The expression of chimeric protein BMP2/EGFP was identified by fluorescent microscope and Western blotting. The bio-activity was examined by the cellular activity and animal heterotopic osteogenesis experiment.
Results: The recombinant plasmid proved successful by restriction enzyme digestion and PCR. The expression of the chimeric protein was shown by fluorescent microscope and Western blotting. The chimeric protein had the double bio-activities of BMP2 and EGFP identified by the cellular activity and animal heterotopic osteogenesis tests.
Conclusion: The recombinant vector of BMP2/pLEGFP is successfully constructed by the gene recombinant technology and its chimeric protein has double bioactivities of BMP2 and EGFP.