关键词: 遗传载体, 基因, 转染

Abstract: To clone the human bone morphogenetic protein-2 (hBMP-2) gene cDNA and construct its replication-deficient type 5 adenoviral vector.Methods Using osteoblastic sarcoma tissue to extract cell total RNA，hBMP-2 cDNA was amplified by RT-PCR.The hBMP-2 cDNA was then inserted into cloning vector pGEM-T.The recombinant plasmid was screened and the recombinant plasmid which was confirmed by restriction enzyme，PCR and sequence analysis was gotten.To construct the recombinant adenoviral vector，the BMP-2 cDNA was subcloned into a shuttle vector and the recombinant pShuttle-BMP2 vector was obtained.It was subcloned into Adeno-X adenoviral genomic DNA and the pAd-BMP2 DNA was obtained. Results A length of 1 200 bp BMP-2 cDNA was cloned，and it was subcloned into adenoviral vector，the recombinant adenovirus was constructed successfully.After measuring the titre of virus（1×10¹¹ nfu•L^-1），the target gene was ev aluated by PCR.Conclusion The hBMP-2 adenoviral is obtained.The successful construction of pAd-BMP2 may provide us useful target for the further research.

Key words: genetic vectors, genes, transfection