Isolation of cDNA clones from an osteosarcoma-ROS17/2.8 library by differential hybridization.
We have used differential hybridization to isolate and characterize two novel cDNAs expressed in chondrocytes and some osteoblastic cells. A rat osteosarcoma ROS17/2.8 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 p10.15 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)2D3, 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.
Waye
M M
MM
Department of Biochemistry, Chinese University of Hong Kong, Shatin, New Territories.
Li
V K
VK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D001859:Bone Neoplasms / Q000235:genetics*; D002356:Cartilage / Q000378:metabolism; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000302:isolation & purification*; D004273:DNA, Neoplasm / Q000302:isolation & purification*; D015723:Gene Library; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000737:chemistry / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D016366:Open Reading Frames; D009928:Organ Specificity; D010006:Osteoblasts / Q000378:metabolism; D012516:Osteosarcoma / Q000235:genetics*; D017433:Protein Structure, Secondary; D051381:Rats; D012886:Skull / Q000166:cytology; D013552:Swine
D018076:DNA, Complementary; D004273:DNA, Neoplasm; D009363:Neoplasm Proteins