Lee C C, Wu X W, Gibbs R A, Cook R G, Muzny D M, Caskey C T
Institute for Molecular Genetics, Baylor College of Medicine, Houston, TX 77030.
Science. 1988 Mar 11;239(4845):1288-91. doi: 10.1126/science.3344434.
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.
尿酸氧化酶（E.C. 1.7.3.3）可催化除人类和某些灵长类动物外的大多数哺乳动物体内尿酸氧化为尿囊素。已确定猪尿酸氧化酶的氨基末端氨基酸序列，并将其用于一种新方法，以生成针对该氨基酸序列的互补DNA（cDNA）探针。该方法基于聚合酶链反应，并利用与氨基酸序列反向翻译产物互补的混合寡核苷酸引物。这种快速简便的cDNA克隆方法具有普遍适用性，仅需部分氨基酸序列。通过该方法开发的cDNA探针用于分离猪尿酸氧化酶全长cDNA，并证明人类中存在同源基因组序列。
