PMID: 16529317Mar 15, 2006Paper

Cloning and constructing of bone morphogenetic protein 2 eukaryotic expression

Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery
Xiaobing TianShu-hua Yang

Abstract

To clone human bone morphogenetic protein 2 (BMP-2) gene and construct the gene's eukaryotic expression vector. 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 then 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 vector was inserted into the pcDNA3. 1(+) eukaryotic expression vector. The agarose electrophoresis showed that the fragments of BMP-2, pGEM-T 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. The pcDNA3. 1 (+)-hBMP-2 eukaryotic vector can be successfully constructed.

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