Polysome immunoprecipitation of phenylalanine hydroxylase mRNA from rat liver and cloning of its cDNA.
The mRNA for phenylalanine hydroxylase (phenylalanine 4-monooxygenase, EC 1.14.16.1) has been purified from total rat liver mRNAs, of which it constitutes less than 0.25%, to greater than 10% purity in a single step by specific polysome immunoprecipitation. The purified mRNA was used for synthesis and cloning of its cDNA. Recombinant colonies containing phenylalanine hydroxylase DNA sequences were identified by differential hybridization, hybrid-selected translation, and blot hybridization analysis. The rat cDNA clone was capable of hybridizing with human phenylalanine hydroxylase mRNA, which will permit the isolation of the corresponding human gene for analysis of phenylketonuria, a hereditary disorder in phenylalanine metabolism that causes permanent mental retardation in humans.
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA; D007158:Immunologic Techniques; D008099:Liver / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D010651:Phenylalanine Hydroxylase / Q000235:genetics* / Q000276:immunology; D010957:Plasmids; D011132:Polyribosomes / Q000032:analysis; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification*; D051381:Rats
D012333:RNA, Messenger; D004247:DNA; D010651:Phenylalanine Hydroxylase