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大鼠肝脏2A型蛋白磷酸酶催化亚基cDNA的分子克隆,以及该基因在正常细胞和癌细胞中高表达的检测。

Molecular cloning of cDNA for the catalytic subunit of rat liver type 2A protein phosphatase, and detection of high levels of expression of the gene in normal and cancer cells.

作者信息

Kitagawa Y, Tahira T, Ikeda I, Kikuchi K, Tsuiki S, Sugimura T, Nagao M

机构信息

Carcinogenesis Division, National Cancer Center Research Institute, Tokyo, Japan.

出版信息

Biochim Biophys Acta. 1988 Nov 10;951(1):123-9. doi: 10.1016/0167-4781(88)90032-2.

Abstract

A cloned cDNA encoding a catalytic subunit of type 2A protein phosphatase from a rat liver cDNA library was obtained by use of a synthetic oligonucleotide corresponding to the tryptic peptide sequence of the purified enzyme. There was only a single amino acid difference between the deduced amino acid sequence of the clone obtained and those of the catalytic subunits, 2A alpha, of the rabbit skeletal muscle, porcine kidney and human liver enzymes, suggesting that this clone was a rat 2A alpha cDNA. On Northern blot analysis using a cDNA fragment as a probe, three mRNA species were detected in rat liver: a major mRNA of 2.0 kb and a minor one of 2.7 kb under high stringency conditions, and also a 1.1 kb mRNA under low stringency conditions. The 2A alpha gene was found to be highly expressed in various tissues of rat, especially the brain. High levels of expression of the gene were also detected in mouse NIH3T3 cells and their transformants, and in human cancer cell lines as well as a human immortalized cell line.

摘要

利用与纯化酶的胰蛋白酶肽段序列相对应的合成寡核苷酸,从大鼠肝脏cDNA文库中获得了一个编码2A型蛋白磷酸酶催化亚基的克隆cDNA。所获得克隆的推导氨基酸序列与兔骨骼肌、猪肾和人肝脏酶的催化亚基2Aα的氨基酸序列之间仅存在一个氨基酸差异,这表明该克隆是大鼠2Aα cDNA。使用cDNA片段作为探针进行Northern印迹分析,在大鼠肝脏中检测到三种mRNA:在高严谨条件下,主要的2.0 kb mRNA和次要的2.7 kb mRNA,以及在低严谨条件下的1.1 kb mRNA。发现2Aα基因在大鼠的各种组织中高度表达,尤其是在大脑中。在小鼠NIH3T3细胞及其转化体、人癌细胞系以及人永生化细胞系中也检测到该基因的高水平表达。

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