人 BMP2和 OPG共表达真核载体的构建与骨质疏松基因治疗相关性的研究

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Objectives To construct human BMP-2 and OPG coexpressing vector and determine its expression in mammalian cells in vitro. Methods Using the isolated total RNA from osteosacoma cell line MG63 as a template,the cDNA encoding hOPG was amplified by reverse transcription polymerase chain reaction (RT PCR) method and cloned into mammalian expression vector pIRES2 EGFP endonucleases sites EcoR I and BamH I,and the BMP-2 encoding region gene was cloned into endonucleases site BstX I.Then the recombinant plasmid pIRES2 OPG BMP2 was transformed into COS 7 cell line,OPG and BMP-2 expression were determined by Western blot assay.Results (1) The sequence of OPG cDNA obtained in this experiment was the same as that of reported, recombinant plasmid pIRES BMP2 OPG was constructed successfully.(2) Human OPG and BMP-2 overexpression cell line COS 7 was selected and confirmed by Western blot analysis. Conclusion Human OPG encoding cDNA,OPG and BMP-2 coexpressing vector and cell lineage expressed OPG and BMP-2 were obtained. These datas suggested coexpressing human OPG and BMP-2 may be a promising and logical apporach for treatment of osteoporosis.
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