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人参 PDR1 样基因在盐胁迫、冷胁迫和激素处理下的分子克隆与表达分析。

Molecular cloning and expression analysis of PDR1-like gene in ginseng subjected to salt and cold stresses or hormonal treatment.

机构信息

Molecular Biology Research Center, School of Life Sciences, Central South University, Changsha 410078, China.

出版信息

Plant Physiol Biochem. 2013 Oct;71:203-11. doi: 10.1016/j.plaphy.2013.07.011. Epub 2013 Jul 31.

Abstract

The plant pleiotropic drug resistance (PDR) family of ATP-binding cassette (ABC) transporters is potentially involved in diverse biological processes. Currently, little is known about their actual physiological functions. A Panax ginseng PDR transporter gene (PgPDR1) was cloned and the cDNA has an open reading frame of 4344 bp. The deduced amino acid sequence contained the characteristic domains of PDR transporters: Walker A, Walker B, and ABC signature. Genomic DNA hybridization analysis indicated that one copy of PgPDR1 gene was present in P. ginseng. Subcellular localization showed that PgPDR1-GFP fusion protein was specifically localized in the cell membrane. Promoter region analysis revealed the presence of cis-acting elements, some of which are putatively involved in response to hormone, light and stress. To understand the functional roles of PgPDR1, we investigated the expression patterns of PgPDR1 in different tissues and under various conditions. Quantitative real-time PCR (qRT-PCR) and Western blotting analysis showed that PgPDR1 was expressed at a high level in the roots and leaves compared to seeds and stems. The expression of PgPDR1 was up-regulated by salicylic acid (SA) or chilling, down-regulated by ABA, and regulated differently at transcript and protein levels by MeJA. These results suggest that PgPDR1 might be involved in responding to environmental stresses and hormones.

摘要

植物多药耐药(PDR)家族的 ATP 结合盒(ABC)转运蛋白可能参与多种生物学过程。目前,人们对它们的实际生理功能知之甚少。本研究克隆了人参 PDR 转运蛋白基因(PgPDR1),cDNA 开放阅读框为 4344bp。推导的氨基酸序列包含 PDR 转运蛋白的特征结构域:Walker A、Walker B 和 ABC 基序。基因组 DNA 杂交分析表明,人参中存在一个 PgPDR1 基因拷贝。亚细胞定位表明 PgPDR1-GFP 融合蛋白特异性定位于细胞膜上。启动子区分析表明存在顺式作用元件,其中一些可能参与对激素、光和胁迫的反应。为了了解 PgPDR1 的功能作用,我们研究了 PgPDR1 在不同组织和不同条件下的表达模式。定量实时 PCR(qRT-PCR)和 Western blotting 分析表明,与种子和茎相比,PgPDR1 在根和叶中表达水平较高。SA 或冷胁迫诱导 PgPDR1 表达上调,ABA 下调 PgPDR1 表达,MeJA 对 PgPDR1 的转录和蛋白水平的调控存在差异。这些结果表明,PgPDR1 可能参与响应环境胁迫和激素。

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