Generation of cDNA probes directed by amino acid sequence: cloning of urate oxidase.

TitleGeneration of cDNA probes directed by amino acid sequence: cloning of urate oxidase.
Publication TypeJournal Article
Year of Publication1988
AuthorsLee, CC, Wu, XW, Gibbs, RA, Cook, RG, Muzny, DM, Caskey, CT
Date Published1988 Mar 11
KeywordsAmino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, DNA, Gene Amplification, Liver, Mice, Molecular Sequence Data, Nucleic Acid Hybridization, Swine, Urate Oxidase

Urate oxidase (E.C. catalyzes the oxidation of uric acid to allantoin in most mammals except humans and certain primates. The amino-terminal amino acid sequence for porcine urate oxidase was determined and used in a novel procedure for generating complementary DNA (cDNA) probes to this amino acid sequence. The procedure is based on the polymerase chain reaction and utilizes mixed oligonucleotide primers complementary to the reverse translation products of an amino acid sequence. This rapid and simple cDNA cloning procedure is generally applicable and requires only a partial amino acid sequence. A cDNA probe developed by this procedure was used to isolate a full-length porcine urate oxidase cDNA and to demonstrate the presence of homologous genomic sequences in humans.

Alternate JournalScience
PubMed ID3344434
Grant ListDK31428 / DK / NIDDK NIH HHS / United States
GM34428 / GM / NIGMS NIH HHS / United States

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