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从胡杨中过表达一种热休克蛋白(ThHSP18.3)赋予酵母应激耐受性。

Overexpression of a heat shock protein (ThHSP18.3) from Tamarix hispida confers stress tolerance to yeast.

机构信息

State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, Harbin, 150040, People's Republic of China.

出版信息

Mol Biol Rep. 2012 Apr;39(4):4889-97. doi: 10.1007/s11033-011-1284-2. Epub 2011 Nov 23.

Abstract

It is well known that plant heat shock proteins (HSPs) play important roles both in response to adverse environmental conditions and in various developmental processes. However, among plant HSPs, the functions of tree plant HSPs are poorly characterized. To improve our understanding of tree HSPs, we cloned and characterized an HSP gene (ThHSP18.3) from Tamarix hispida. Sequence alignment reveals that ThHSP18.3 belongs to the class I small heat shock protein family. A transient expression assay showed that ThHSP18.3 protein was targeted to the cell nucleus. Treatment of Tamarix hispida with cold and heat shock highly induced ThHSP18.3 expression in all studied leaves, roots and stems, whereas, treatment of T. hispida with NaCl, NaHCO(3), and PEG induced ThHSP18.3 expression in leaves and decreased its expression in roots and stems. Further, to study the role of ThHSP18.3 in stress tolerance under different stress conditions, we cloned ThHSP18.3 into the pYES2 vector, transformed and expressed the vector in yeast Saccharomyces cerevisiae. Yeast cells transformed with an empty pYES2 vector were employed as a control. Compared to the control, yeast cells expressing ThHSP18.3 showed greater tolerance to salt, drought, heavy metals, and both low and high temperatures, indicating that ThHSP18.3 confers tolerance to these stress conditions. These results suggested that ThHSP18.3 is involved in tolerance to a variety of stress conditions in T. hispida.

摘要

众所周知,植物热休克蛋白(HSPs)在应对不利环境条件和各种发育过程中都发挥着重要作用。然而,在植物 HSPs 中,树木植物 HSPs 的功能尚未得到很好的描述。为了提高我们对树木 HSPs 的理解,我们从柽柳中克隆并鉴定了一个 HSP 基因(ThHSP18.3)。序列比对表明,ThHSP18.3 属于 I 类小热休克蛋白家族。瞬时表达分析表明,ThHSP18.3 蛋白靶向细胞核。对柽柳进行冷和热休克处理,在所有研究的叶片、根和茎中高度诱导 ThHSP18.3 的表达,而用 NaCl、NaHCO(3) 和 PEG 处理柽柳,在叶片中诱导 ThHSP18.3 的表达,而在根和茎中降低其表达。此外,为了研究 ThHSP18.3 在不同胁迫条件下对胁迫耐受的作用,我们将 ThHSP18.3 克隆到 pYES2 载体中,转化并在酵母酿酒酵母中表达该载体。将空 pYES2 载体转化的酵母细胞用作对照。与对照相比,表达 ThHSP18.3 的酵母细胞对盐、干旱、重金属以及低温和高温表现出更强的耐受性,表明 ThHSP18.3 赋予了对这些胁迫条件的耐受性。这些结果表明,ThHSP18.3 参与了柽柳对多种胁迫条件的耐受。

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