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豌豆中一种丝裂原活化蛋白激酶同源物的分子克隆与表达

Molecular cloning and expression of a MAP kinase homologue from pea.

作者信息

Stafstrom J P, Altschuler M, Anderson D H

机构信息

Plant Molecular Biology Center, Northern Illinois University, DeKalb 60115.

出版信息

Plant Mol Biol. 1993 Apr;22(1):83-90. doi: 10.1007/BF00038997.

Abstract

The cdc2 kinases are important cell cycle regulators in all eukaryotes. MAP kinases, a closely related family of protein kinases, are involved in cell cycle regulation in yeasts and vertebrates, but previously have not been documented in plants. We used PCR to amplify Brassica napus DNA sequences using primers corresponding to amino sequences that are common to all known protein kinases. One sequence was highly similar to KSS1, a MAP kinase from Saccharomyces cerevisiae. This sequence was used to isolate a full-length MAP kinase-like clone from a pea cDNA library. The pea clone, called D5, shared approximately 50% amino acid identity with MAP kinases from yeasts and vertebrates and about 41% identity with plant cdc2 kinases. An expression protein encoded by D5 was recognized by an antiserum specific to human MAP kinases (ERKs). Messenger RNA corresponding to D5 was present at similar levels in all tissues examined, without regard to whether cell division or elongation were occurring in those tissues.

摘要

cdc2激酶是所有真核生物中重要的细胞周期调节因子。丝裂原活化蛋白激酶(MAP激酶)是与之密切相关的一类蛋白激酶,参与酵母和脊椎动物的细胞周期调控,但此前在植物中尚未有相关记载。我们使用PCR,以对应于所有已知蛋白激酶共有氨基酸序列的引物扩增甘蓝型油菜的DNA序列。其中一个序列与酿酒酵母的MAP激酶KSS1高度相似。利用该序列从豌豆cDNA文库中分离出一个全长MAP激酶样克隆。这个名为D5的豌豆克隆与酵母和脊椎动物的MAP激酶约有50%的氨基酸同一性,与植物cdc2激酶约有41%的同一性。D5编码的表达蛋白能被人MAP激酶(细胞外信号调节激酶)特异性抗血清识别。对应于D5的信使核糖核酸在所有检测的组织中含量相似,无论这些组织中是否发生细胞分裂或细胞伸长。

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