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[谷胱甘肽S-转移酶基因过表达促进盐胁迫下转基因拟南芥的生长]

[Overexpression of GST gene accelerates the growth of transgenic Arabidopsis under salt stress].

作者信息

Qi Yuan-Cheng, Zhang Shi-Min, Wang Li-Ping, Wang Ming-Dao, Zhang Hui

机构信息

College of Biological Technique and Food Science, Henan Agricultural University, Zhengzhou 450002, China.

出版信息

Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2004 Oct;30(5):517-22.

Abstract

The Suaeda salsa glutathione s-transferase gene (GST) was inserted downstream of the 35S promoter in the plant expression vector pROK II and then was introduced into Arabidopsis thaliana by Agrobacterium tumefaciens through floral dip method. Transformants were selected for their ability to grow on medium containing kanamycin. The fact that the GST gene had been transferred into the Arabidopsis thaliana genome was confirmed by the PCR-Southern blotting analysis. After cultivation, independent homozygous transgenic lines were obtained after selection of T(3) progenies on MS medium containing kanamycin. The expression of the gene transferred into the Arabidopsis thaliana was confirmed by Northern blotting. During salt stress, analysis of total glutathione (both oxidized and reduced type) and biomass of transgenic and wild Arabidopsis. The biomass of transgenic lines (GT) was slightly but significantly greater than that of wild type line (WT), and levels of oxidized glutathione (GSSG) were significantly higher in transgenic lines than in wild type. Therefore, overexpression of GST can increase Arabidopsis growth under salt stress, and this effect can be caused by oxidation of the reduced glutathione (GSH ).

摘要

将盐地碱蓬谷胱甘肽 - S - 转移酶基因(GST)插入植物表达载体pROK II的35S启动子下游,然后通过农杆菌介导的浸花法将其导入拟南芥。通过筛选在含有卡那霉素的培养基上生长的能力来选择转化体。通过PCR - Southern印迹分析证实GST基因已转入拟南芥基因组。培养后,在含有卡那霉素的MS培养基上选择T(3)代后代后获得独立的纯合转基因株系。通过Northern印迹证实转入拟南芥的基因的表达。在盐胁迫期间,分析转基因和野生拟南芥的总谷胱甘肽(氧化型和还原型)和生物量。转基因株系(GT)的生物量略高于但显著高于野生型株系(WT),并且转基因株系中氧化型谷胱甘肽(GSSG)的水平显著高于野生型。因此,GST的过表达可以增加盐胁迫下拟南芥的生长,并且这种效应可能是由还原型谷胱甘肽(GSH)的氧化引起的。

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