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拟南芥中一个假定的果胶甲酯酶cDNA的分子克隆与特性分析

Molecular cloning and characterisation of a putative pectin methylesterase cDNA in Arabidopsis thaliana (L.).

作者信息

Richard L, Qin L X, Gadal P, Goldberg R

机构信息

Institut Jacques Monod, Paris, France.

出版信息

FEBS Lett. 1994 Nov 28;355(2):135-9. doi: 10.1016/0014-5793(94)01187-7.

Abstract

Pectin methylesterase (PME) is a cell wall enzyme that catalyses the de-esterification of pectins leading to fundamental changes which confer new properties to the micro-environment of each cell. In order to elucidate the meaning of PME-mediated changes of pectin in the time course of cell differentiation, we attempted to study the regulation of PME genes in Arabidopsis thaliana. In this report, the first full cDNA sequence showing sequence similarities with other PME genes characterised so far in other plant species has been isolated from an Arabidopsis shoot cDNA library. This ATPMEl cDNA is 1,970 bp long and contains an open reading frame encoding a protein of 64.1 kDa and a basic pI of 8.7 as predicted from the nucleotide sequence. Northern blot analyses denoted changes in the expression level of the ATPMEl mRNA according to plant organs. High mRNA levels were found in young developing organs such as cauline leaves while they were significantly lower in rosette leaves, stems and inflorescences, and almost undetectable in roots. Beside this molecular approach, isoelectrofocusing analyses revealed the occurrence of three PME isoforms in Arabidopsis. Two PME isoforms with pI values of 4.9 and 9.1 were found throughout the plant, but at a higher level in the root, while an other PME isoform with a pI of 5.7 was essentially detected in the inflorescence. The relationship between our observations and the data reported for other plant species is discussed.

摘要

果胶甲基酯酶(PME)是一种细胞壁酶,它催化果胶的去酯化反应,导致果胶发生根本性变化,从而赋予每个细胞微环境新的特性。为了阐明在细胞分化过程中PME介导的果胶变化的意义,我们试图研究拟南芥中PME基因的调控。在本报告中,从拟南芥茎尖cDNA文库中分离出了第一个与迄今在其他植物物种中鉴定的其他PME基因具有序列相似性的完整cDNA序列。这个ATPMEl cDNA长1970 bp,包含一个开放阅读框,根据核苷酸序列预测,该开放阅读框编码一个64.1 kDa的蛋白质,碱性pI为8.7。Northern印迹分析表明,ATPMEl mRNA的表达水平因植物器官而异。在幼嫩的发育器官如茎生叶中发现了较高的mRNA水平,而在莲座叶、茎和花序中则显著较低,在根中几乎检测不到。除了这种分子方法外,等电聚焦分析还揭示了拟南芥中存在三种PME同工型。在整个植物中发现了两种pI值分别为4.9和9.1的PME同工型,但在根中的含量较高,而另一种pI为5.7的PME同工型主要在花序中检测到。我们讨论了我们的观察结果与其他植物物种报道的数据之间的关系。

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