title: Isolation of cDNA clones coding for the catalytic subunit of mouse cAMP-dependent protein kinase.
abstract: mRNA coding for the catalytic (C) subunit of cAMP-dependent protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37) was partially purified from bovine testis by polysome immunoadsorption and oligo(dT)-chromatography. This enriched mRNA preparation was used to prepare and differentially screen a cDNA library. One of the selected cDNA clones was shown to hybrid-select mRNA coding for a 40-kDa protein that was specifically precipitated with antibodies to the C subunit. This bovine cDNA clone was then used to isolate a series of mouse cDNA clones that are complementary to the entire mouse C subunit mRNA. The mouse clones code for a protein of 351 amino acids that shows 98% homology to the bovine C subunit and hybridize to a single mRNA of 2.4 kilobases in mouse heart and brain. Southern blot analysis of total genomic DNA suggests that there is a single mouse gene coding for the C subunit. mRNA levels for both the C subunit and the type I regulatory subunit in various mouse tissues and cell lines were quantitated and compared by using single-stranded RNA probes prepared with SP6 polymerase.
authors.forename: M D
authors.initials: MD
authors.lastname: Uhler
authors.forename: D F
authors.initials: DF
authors.lastname: Carmichael
authors.forename: D C
authors.initials: DC
authors.lastname: Lee
authors.forename: J C
authors.initials: JC
authors.lastname: Chrivia
authors.forename: E G
authors.initials: EG
authors.lastname: Krebs
authors.forename: G S
authors.initials: GS
authors.lastname: McKnight
mesh_terms: D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005786:Gene Expression Regulation; D051379:Mice; D011494:Protein Kinases / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D014018:Tissue Distribution
chemical_list: D012333:RNA, Messenger; D004247:DNA; D011494:Protein Kinases