| 摘要 |
PURPOSE:To obtain a new cDNA, having a specific base sequence, coding for a receptor type tyrosine kinase part related to a human osteoblast and useful for developing, etc., a medicine or a diagnostic agent concerned in the bone metabolism causing the growth, differentiation and activation of the osteoblast. CONSTITUTION:This new cDNA capable of coding an enzymic part is obtained by culturing and subculturing a human osteoblastic cell strain MG63 in a culture medium containing 10% bovine fetal blood serum under conditions of 37 deg.C in a CO2 incubator, extracting the whole RNA from the recovered cell according to a lithium chloride urea method, isolating an mRNA with an oligodT column, then synthesizing a cDNA using the mRNA as a template, amplifying the prepared cDNA with a mixed primer which is a subdomain of a tyrosine kinase and has a recognition site for a restriction enzyme connected thereto according to a genetic polymerase chain reaction (PCR) and cloning the resultant PCR product. Thereby, the resultant DNA has a base sequence expressed by the formula and is effective in developing a medicine or a diagnostic agent concerned in the bone metabolism causing the growth, differentiation and activation of an osteoblast and providing the whole length gene of this enzyme. |
