Objective To clone human bone morphogenetic protein 2 ( BMP-2) gene and construct the gene’s eukaryotic expression vector. Methods The total RNA was extracted from human osteosarcoma cells, the human BMP-2 cDNA was amplified by RT-PCR and inserted into pGEM-T vector. The positive clones were screened out, and the n the recombinant plasmid was confirmed by restriction enzyme digestion, PCR and the analysis of nucleotide sequence. The BMP-2 cDNA in the pGEM-T cloning vec tor was inserted into the pcDNA3.1(+) eukaryotic expression vector. Results The agarose electrophoresis showed that the fragments of BMP-2, pGEMT and pcDNA3.1(+) were 1.2 kbp, 4.0 kbp and 5.0 kbp, respectively. The result of nucleotide sequence confirmed that the cDNA sequence, which was inserted into pGEM-T and pcDNA3.1(+) plasmid was human BMP-2. Conclusion The pcDNA3.1(+)-hBMP-2 eukaryotic vector can be successfully constructed.
Citation： TIAN Xiaobing,SUN Li,YANG Shuhua.. CLONING AND CONSTRUCTING OF BONE MORPHOGENETIC PROTEIN 2 EUKARYOTIC EXPERSSION VECTOR. Chinese Journal of Reparative and Reconstructive Surgery, 2006, 20(2): 112-115. doi: Copy