A study on growth factors regulating bone formation

Part 1 cDNA cloning of murine bone morphogenetic protein by polymerase chain reaction

Abstract

A cDNA clone of murine bone morphogenetic protein (BMP) was obtained by polymerase chain reaction, using oligonucleotides as primers derived from human BMP2 and BMP4. Messenger RNAs were isolated from neonatal murine tissue and used for the synthesis of cDNA with reverse transcriptase.
Specifically amplified cDNAs were cloned into bluescript plasmid and subjected to sequencing. One clone which partially encoded murine BMP2 was obtained. The nucleotide sequence, 554 bp in length, shared 88.6% identity with that of human BMP2 and the deduced amino acids including the mature, biologically active protein exhibited 96.2% homology. Within the C-terminal 114 amino acids of the protein, seven cysteine residues were completely conserved which is characteristic of TGF-β superfamily. Substitution of only three amino acids was found when compared with the corresponding human sequence. Isolation of the full length cDNA is now in progress.