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Isolation of cDNA clones from an osteosarcoma‐ROS17/2.8 library by differential hybridization
Author(s) -
Waye Mary M. Y.,
Li Vincent K. C.
Publication year - 1994
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240540303
Subject(s) - complementary dna , microbiology and biotechnology , cdna library , biology , calvaria , population , clone (java method) , messenger rna , rna , dna , gene , genetics , in vitro , demography , sociology
Abstract 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) 2 D 3 , 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.