Waye M M, Li V K
Department of Biochemistry, Chinese University of Hong Kong, Shatin, New Territories.
J Cell Biochem. 1994 Mar;54(3):273-80. doi: 10.1002/jcb.240540303.
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.
我们利用差异杂交技术分离并鉴定了在软骨细胞和一些成骨细胞中表达的两个新的cDNA。对大鼠骨肉瘤ROS17/2.8 cDNA文库进行筛选，分离出与放射性标记的猪颅骨cDNA强烈杂交但与对照放射性标记cDNA弱杂交的cDNA克隆。获得了两个克隆——p.6.1和p.10.15。结果显示，p10.15的放射性标记探针能与来自大鼠颅骨-RCJ 3.1C5.18的软骨生成克隆细胞群体的2.3 Kb信使RNA特异性杂交，并且该信使RNA被抑制这些细胞软骨生成的1,25(OH)2D3下调。另一个克隆p6.1能与一个0.95 Kb的信使RNA杂交，该信使RNA在大鼠肝脏、肾脏、肺、肌肉和大脑中表达，但在脾脏中不表达，在胸腺中仅低水平表达。
