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一项转录组学分析,用于鉴定将异源抗草甘膦5-烯醇丙酮酰莽草酸-3-磷酸合酶引入大肠杆菌的意外影响。

A transcriptomic analysis for identifying the unintended effects of introducing a heterologous glyphosate-tolerant EPSP synthase into Escherichia coli.

作者信息

Li Liang, Zhou Zhengfu, Jin Wujun, Wan Yusong, Lu Wei

机构信息

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China.

出版信息

Mol Biosyst. 2015 Mar;11(3):852-8. doi: 10.1039/c4mb00566j. Epub 2015 Jan 7.

Abstract

Glyphosate is one of the most commonly used broad-spectrum herbicides with little to no hazard to animals, human beings, or the environment. Some microbial 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase variants are not inhibited by glyphosate, and they provide a powerful tool to engineer glyphosate-tolerant plants. However, the unintended effects of EPSP synthase expression patterns on microbes are not yet clear. Here, we use an Affymetrix GeneChip analysis to study how introduction of a heterologous glyphosate-tolerant EPSP synthase into a model microorganism Escherichia coli (E. coli) affects the global gene expression profile. The profile showed that 161 of 4071 genes were differentially expressed after the introduction of the synthase: 19 (0.47%) were up-regulated and 143 (3.49%) were down-regulated. The microarray results, in combination with BiOLOG substrate utilization and amino acid composition assays, suggested that heterologous EPSP synthase expression had very minor effects on E. coli. Although a small number of genes and metabolites were affected by EPSP synthase expression, no functional correlations were identified among the dataset. This study may shed light on the effect of EPSP synthase expression on microbes, which should help in the assessment of environmental safety.

摘要

草甘膦是最常用的广谱除草剂之一,对动物、人类或环境几乎没有危害。一些微生物的5-烯醇丙酮酸莽草酸-3-磷酸(EPSP)合酶变体不受草甘膦抑制,它们为培育耐草甘膦植物提供了有力工具。然而,EPSP合酶表达模式对微生物的非预期影响尚不清楚。在此,我们使用Affymetrix基因芯片分析来研究将异源耐草甘膦EPSP合酶引入模式微生物大肠杆菌后如何影响全局基因表达谱。该谱显示,引入合酶后,4071个基因中有161个差异表达:19个(0.47%)上调,143个(3.49%)下调。微阵列结果与BiOLOG底物利用和氨基酸组成分析相结合表明,异源EPSP合酶表达对大肠杆菌的影响非常小。虽然少数基因和代谢物受EPSP合酶表达影响,但数据集中未发现功能相关性。本研究可能有助于揭示EPSP合酶表达对微生物的影响,这应有助于环境安全性评估。

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