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PubMed · 16529317

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

Xiaobing Tian·Li Sun·Shuhua Yang
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Abstract

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. CONCLUSION: The pcDNA3. 1 (+)-hBMP-2 eukaryotic vector can be successfully constructed.

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Bone Morphogenetic Protein 2Cell Line, TumorCloning, MolecularDNA, ComplementaryDNA, RecombinantGene ExpressionGenetic VectorsHumansPlasmidsReverse Transcriptase Polymerase Chain ReactionTransfectionEnglish AbstractJournal ArticleResearch Support, Non-U.S. Gov't

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Xiaobing Tian, Li Sun, Shuhua Yang. 2006. [Cloning and constructing of bone morphogenetic protein 2 eukaryotic expression].. https://pubmed.ncbi.nlm.nih.gov/16529317/

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