New expression of bone morphogenetic protein 2 adenoviral vector eukaryotic cells and identification of.docVIP

 PAGE \* MERGEFORMAT 19 New expression of bone morphogenetic protein 2 adenoviral vector eukaryotic cells and identification of Of: Zhang Zheng, Liu Danping, Chenjun Jiang, Guo Tao, Zhang M [Abstract] Objective To construct the expression of both FLAG epitope tag of bone morphogenetic protein 2 (BMP2) and reports of target protein molecules of green fluorescent protein (GFP) adenovirus new eukaryotic expression vector. Methods to remove translation stop codon child and add a new restriction enzyme recognition sites in the BMP2 gene targeting Xho connected to the adenovirus shuttle plasmid pShuttle CMV-IRES-hrGFP-1. BMP2 + recombinant adenovirus carrying the shuttle plasmid by restriction endonuclease, Pme digested linearized into E. coli BJ5183-AD-1, using homologous recombination mechanism BMP2 + and GFP gene expression of components together with their homeopathic recombinant plasmid into the adenovirus genome. digested with Pac remove the plasmid composition, exposure to adenovirus genome reverse terminal repeat sequence, transfected into 293 cells, the adenovirus packaging, complete a new adenovirus vector Ad CMV-BMP2 +-IRES-GFP-1 Construction. as a positive control, while widely used for years prepared the adenovirus vector BMP2 Ad CMV-BMP2. vector infection in two marrow stromal cells by RT-PCR and fluorescent microscopy were used to detect bone morphogenetic protein-2 and green fluorescent protein; by alkaline phosphatase activity assay to determine the role of BMP2 induced osteogenic differentiation. Results mutated bone morphogenetic protein 2 gene sequence, recombinant adenovirus shuttle plasmid and the plasmid restriction map to meet the intended design. infection after bone marrow stromal cells, while a new expression of adenovirus vector can induce reporter molecule GFP and osteogenic differentiation activity of BMP2. BMP2 expression is successfully constructed a new adenovirus vector. [Keywords:] bone morphogenetic protein 2 gene; gree