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一种玉米花粉特异性钙依赖且不依赖钙调蛋白的蛋白激酶的克隆与特性分析

Cloning and characterization of a maize pollen-specific calcium-dependent calmodulin-independent protein kinase.

作者信息

Estruch J J, Kadwell S, Merlin E, Crossland L

机构信息

Department of Plant Molecular Biology, Ciba Agricultural Biotechnology, Research Triangle Park, NC 27709.

出版信息

Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8837-41. doi: 10.1073/pnas.91.19.8837.

Abstract

A calcium-dependent calmodulin-independent protein kinase (CDPK) has been cloned from maize (Zea mays). The sequence predicts a 550-amino acid (predicted molecular mass is 60 kDa) protein with two major functional domains: an N-terminal catalytic domain highly homologous to protein kinases and a C-terminal domain resembling calmodulins. Northern analysis shows that the expression of the maize CDPK gene is pollen specific and that its transcription is restricted to late stages of pollen development. Western blots reveal a major abundance of CDPK protein at the stage of pollen germination. In vitro germination and pollen tube growth are impaired upon addition of a calmodulin antagonist (calmidazolium), CDPK inhibitors (W-7), and antisense oligonucleotides directed against CDPK mRNA. These observations indicate that the function of the pollen-specific maize CDPK protein is required for germination and pollen tube growth.

摘要

一种钙依赖性、不依赖钙调蛋白的蛋白激酶(CDPK)已从玉米(Zea mays)中克隆出来。该序列预测有一个550个氨基酸(预测分子量为60 kDa)的蛋白,具有两个主要功能结构域:一个与蛋白激酶高度同源的N端催化结构域和一个类似于钙调蛋白的C端结构域。Northern分析表明,玉米CDPK基因的表达具有花粉特异性,其转录仅限于花粉发育的后期阶段。Western印迹显示在花粉萌发阶段CDPK蛋白大量存在。添加钙调蛋白拮抗剂(氯丙咪嗪)、CDPK抑制剂(W-7)以及针对CDPK mRNA的反义寡核苷酸后,体外萌发和花粉管生长受到损害。这些观察结果表明,花粉特异性玉米CDPK蛋白的功能是萌发和花粉管生长所必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bbd3/44701/89ff161bd761/pnas01141-0120-a.jpg

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