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Science 11 March 1988:
Vol. 239. no. 4845, pp. 1288 - 1291
DOI: 10.1126/science.3344434
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Articles

Science, Vol 239, Issue 4845, 1288-1291
Copyright © 1988 by American Association for the Advancement of Science

articles

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

CC Lee, XW Wu, RA Gibbs, RG Cook, DM Muzny, and CT Caskey

Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.

Urate oxidase (E.C. 1.7.3.3) 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.


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