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水稻中谷胱甘肽S-转移酶基因的克隆、分子特征分析及异源表达

Cloning, molecular characterization and heterologous expression of a glutathione S-transferase gene in rice.

作者信息

Hu T Z, He S, Huang X Y, Deng L, Wang G X

机构信息

Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing 400044, China.

出版信息

Bioorg Khim. 2011 May-Jun;37(3):386-92. doi: 10.1134/s1068162011030174.

Abstract

OsGSTL2 is one of three tandem-arranged glutathione S-transferase, lambda class genes in chromosome 3 of rice (Oryza sativa L.). It includes 9 introns and 10 exons, and encodes a protein of 244 amino acid residues with a calculated molecular mass of 27.37 kDa. The predicted three-dimensional structure of OsGSTL2 showed a typical glutathione S-transferase fold. Using semi-quantitative RT-PCR analysis, OsGSTL2 transcript was detected in the roots and leaves of seedling stage and tillering stage, and the roots, leaves and panicles of heading stage from rice plants, and the expression level of OsGSTL2 mRNA in rice roots show significant change under chlorsulfuron stress. The OsGSTL2 gene was cloned into pYTV vector and was transformed into yeast strain PEP4. Western blot analysis showed the exogenous OsGSTL2 was expressed in transformed yeast. GST activity of crude extracts of yeast showed the OsGSTL2 transgenic yeast had higher levels of GST activities than control yeasts. These findings suggested that the OsGSTL2 is a glutathione S-transferase and has potential use in detoxification.

摘要

OsGSTL2是水稻(Oryza sativa L.)第3号染色体上3个串联排列的谷胱甘肽S-转移酶λ类基因之一。它包含9个内含子和10个外显子,编码一个由244个氨基酸残基组成的蛋白质,计算分子量为27.37 kDa。预测的OsGSTL2三维结构显示出典型的谷胱甘肽S-转移酶折叠。通过半定量RT-PCR分析,在水稻幼苗期和分蘖期的根和叶、抽穗期的根、叶和穗中检测到OsGSTL2转录本,并且在氯磺隆胁迫下水稻根中OsGSTL2 mRNA的表达水平发生显著变化。将OsGSTL2基因克隆到pYTV载体中并转化到酵母菌株PEP4中。蛋白质免疫印迹分析表明外源OsGSTL2在转化酵母中表达。酵母粗提物的GST活性表明,OsGSTL2转基因酵母的GST活性水平高于对照酵母。这些发现表明OsGSTL2是一种谷胱甘肽S-转移酶,在解毒方面具有潜在用途。

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