Construction and identification of human BMP-7 gene recombinant adenovirus

ObjectiveTo construct and identify the recombinant adenovirus encoding human BMP-7 gene and observe its ability to infect HKC cells.MethodsThe human BMP-7 gene fragment was cloned into the shuttle plasmid pDC316 to form pDC316-BMP-7.Then pDC316-BMP-7 and the framework plasmid pBHGlox_E1,3Cre were transfected into HEK293 cells for homogenous recombination in cells with Lipofectamine 2000.The target gene was detected by polymerase chain reaction(PCR).The titer and its infection rate were determined using the green fluorescent protein(GFP)expression in the shuttle plasmid.ResultsRestriction endonuclease and PCR analysis confirmed that the human BMP-7 gene was successfully inserted into the adenovirus vector.The titer of the recombinant adenovirus was 1.25×1010 PFU/ml.The adenovirus has a strong effect on human HKC cells.ConclusionThe recombinant adenovirus containing human BMP-7 gene was successfully constructed by the method of homogenous recombination in cells.