[Cloning and constructing of bone morphogenetic protein 2 eukaryotic expression].
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 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.


RESULTS
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.


CONCLUSIONS
The pcDNA3. 1 (+)-hBMP-2 eukaryotic vector can be successfully constructed.
Tian
Xiaobing
X
Department of orthopedics, People's Hospital of Guizhou Province, Guiyang Guizhou 550002, P. R. China. txb6@vip.163.com
Sun
Li
L
Yang
Shuhua
S
D055396:Bone Morphogenetic Protein 2 / Q000235:genetics*; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004274:DNA, Recombinant; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D055396:Bone Morphogenetic Protein 2; D018076:DNA, Complementary; D004274:DNA, Recombinant