Isolation of cDNA clones from an osteosarcoma-ROS17/2.8 library by differential hybridization

✍ Scribed by Dr. Mary M. Y. Waye; Vincent K. C. Li

• Book ID: 102876580
• Publisher: John Wiley and Sons
• Year: 1994
• Tongue: English
• Weight: 678 KB
• Volume: 54
• Category: Article
• ISSN: 0730-2312
• DOI: 10.1002/jcb.240540303

✦ Synopsis

We have used differential hybridization to isolate and characterize two novel cDNAs expressed in chondrocytes and some osteoblastic cells. A rat osteosarcoma ROSI 7/2.8 cDNA library was screened and cDNA clones hybridizing strongly to radiolabeled porcine calvaria cDNA but weakly to a control radiolabeled cDNA were isolated. Two clones were obtained-p.6.1 and p.10.15. A radiolabeled probe of pl0.15 was shown to hybridize specifically to a 2.3 Kb message RNA from a chondrogenic clonal cell population from rat calvaria-RCJ 3.1C5.18, and the mRNA was downregulated by 1,25 (OH)>D3, which inhibits chondrogenesis in these cells. The other clone, p6.1, was found to hybridize to a 0.95 Kb message that is expressed in rat liver, kidney, lung, muscle, and brain, but not expressed in spleen and expressed only in low levels in thymus.