CLONING OF A CDNA FOR A MAJOR HUMAN PROTEIN-TYROSINE-PHOSPHATASE
CLONING OF A CDNA FOR A MAJOR HUMAN PROTEIN-TYROSINE-PHOSPHATASE
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DOI：
10.1073/pnas.87.7.2735
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发表时间：
1990-04-01
影响因子：
11.1
通讯作者：
NEEL, BG
中科院分区：
文献类型：
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作者：
CHERNOFF, J;SCHIEVELLA, AR;NEEL, BG
我们分离了一个编码人胎盘主要蛋白酪氨酸磷酸酶（Protein-tyrosine-phosphate phosphohydrolase，EC 3.1.3.48）的cDNA克隆。简并寡核苷酸，基于蛋白质的氨基酸序列，用于通过聚合酶链反应从人胎盘cDNA扩增基因的内部片段。然后将该片段用于探测人胎盘cDNA文库。分离3.3-内切酶（kb）插入片段并测序。该插入片段具有一个单一的延伸开放阅读框，预测Mr. apprxeq的435个氨基酸的蛋白质。50,000.从氨基末端到残基321，推导的氨基酸序列与先前通过肽测序确定的氨基酸序列相同[Charbonneau，H.，Tonks，N. K.，库马尔，S.，迪尔茨角D. Harrylock，M.，酷D E. Krebs，E. G.，Fischer，E. H.和沃尔什，K. A.等人（1989）Proc. Acad. Sci. USA 86，5252-5256];然而，序列预测蛋白质含有在报道的肽序列中不存在的另外114个氨基酸。在体外翻译的3.3 kb插入产生的蛋白质的Mr 56,000，在一般协议与预测的大小。磷酸酶基因似乎在人类基因组DNA中以单拷贝存在，并在多种组织中转录成3.5 kb的信息。
We have isolated a cDNA clone encoding the major protein-tyrosine-phosphatase (protein-tyrosine-phosphate phosphohydrolase, EC 3.1.3.48) of human placenta. Degenerate oligonucleotides, based on the amino acid sequence of the protein, were used to amplify an internal fragment of the gene from human placental cDNA by the polymerase chain reaction. This fragment was then used to probe a human placental cDNA library. A 3.3-kilobase (kb) insert was isolated and sequenced. The insert has a single extended open reading frame that predicts a 435 amino acid protein of Mr .apprxeq. 50,000. From the amino terminus to residue 321, the deduced amino acid sequence is identical to that previously determined by peptide sequencing [Charbonneau, H., Tonks, N. K., Kumar, S., Diltz, C. D. Harrylock, M., Cool, D. E. Krebs, E. G., Fischer, E. H. and Walsh, K. A. (1989) Proc. Natl. Acad. Sci. USA 86, 5252-5256]; however, the sequence predicts that the protein contains an additional 114 amino acids not present in the reported peptide sequence. In vitro translation of the 3.3-kb insert produces a protein of Mr 56,000, in general agreement with the predicted size. The phosphatase gene appears to present as a single copy in human genomic DNA and is transcribed into a 3.5-kb message in a variety of tissues.