Cloning and molecular analyses of the Arabidopsis thaliana chloride channel gene family
Cloning and molecular analyses of the Arabidopsis thaliana chloride channel gene family
复制标题
拟南芥氯通道基因家族的克隆和分子分析
DOI：
10.1016/j.plantsci.2009.02.006
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发表时间：
2009-05-01
期刊：
影响因子：
5.2
通讯作者：
Zhang, Hong-xia
中科院分区：
文献类型：
--
作者：
Lv, Qun-dan;Tang, Ren-jie;Zhang, Hong-xia
拟南芥氯通道（CLC）家族包括7个成员，AtCLCa-g，可分为两个不同的亚类。本研究展示了拟南芥CLC （AtCLC）基因家族的发育和组织特异性表达，以及个体成员的亚细胞定位。反转录PCR （RT-PCR）分析表明，这些家族成员在拟南芥中普遍存在，但存在差异表达。启动子驱动的β -葡萄糖醛酸酶（GUS）在转基因拟南芥植物中的表达进一步证实了该家族重叠但不同的表达模式，所有家族成员都主要在维管组织中表达，AtCLCc在保护细胞中优先表达。当AtCLCe在烟草叶肉原生质体中以黄色荧光蛋白（YFP）标记的融合蛋白表达时，AtCLCe特异性定位于叶绿体；AtCLCa-c和g定位于液泡膜，而AtCLCd和AtCLCf定位于高尔基体。在7个AtCLC成员中，只有AtCLCc和AtCLCd可以抑制缺乏单氯通道gef1的酵母突变体Delta gef1的阳离子敏感表型，但效果不同。2009爱思唯尔爱尔兰有限公司版权所有。
The Arabidopsis chloride channels (CLC) family includes seven members, AtCLCa-g, that can be divided into two distinct subclasses. Here we show the developmental and tissue-specific expression of Arabidopsis CLC (AtCLC) gene family, and the subcellular localization of individual members. Reverse transcription PCR (RT-PCR) analyses indicated that the family members were ubiquitously but differentially expressed in Arabidopsis. Promoter-driven beta-glucuronidase (GUS) expression in transgenic Arabidopsis plants further demonstrated the overlapping but distinct expression pattern of this family, and all the family members were predominantly expressed in vascular tissues, as well as a preferential expression of AtCLCc in guard cells. When expressed as yellow fluorescent protein (YFP)tagged fusion proteins in tobacco mesophyll protoplasts, AtCLCe was specifically localized to chloroplast; AtCLCa-c and g were localized to the vacuole membranes, whereas AtCLCd and AtCLCf were targeted to Golgi apparatus. Among the seven AtCLC members, only AtCLCc and AtCLCd could suppress, with discrepant efficacy, the cation-sensitive phenotype of Delta gef1, a yeast mutant lacking the single chloride channel GEF1. (C) 2009 Elsevier Ireland Ltd. All rights reserved.