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互补筛选、鉴定新型枯草芽孢杆菌 II 型 5-烯醇式丙酮酰莽草酸-3-磷酸合酶并加以应用。

Complementary screening, identification and application of a novel class II 5-enopyruvylshikimate-3-phosphate synthase from Bacillus cereus.

机构信息

Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, People's Republic of China.

出版信息

World J Microbiol Biotechnol. 2013 Mar;29(3):549-57. doi: 10.1007/s11274-012-1209-9. Epub 2012 Nov 18.

Abstract

A novel aroA gene encoding 5-enolpyruvylshikimate-3-phosphate synthase from Bacillus cereus was identified and overexpressed by genomic library construction and complementary screening. The enzyme was then purified to homogeneity. We also transformed the aroA ( B. cereus ) gene into Arabidopsis thaliana by a floral dip method, and demonstrated that transgenic A. thaliana plants exhibited significant glyphosate resistance compared with the wild type. These results strongly suggested that the strategy was highly efficient and advantageous for rapidly cloning aroA genes from microorganisms in natural environments.

摘要

从蜡状芽孢杆菌中鉴定并通过基因组文库构建和互补筛选过表达了一个新型编码 5-烯醇丙酮酰莽草酸-3-磷酸合酶的aroA 基因。然后,该酶被纯化至均一状态。我们还通过花浸法将aroA(蜡状芽孢杆菌)基因转入拟南芥,结果表明与野生型相比,转基因拟南芥植物表现出明显的草甘膦抗性。这些结果强烈表明,该策略对于从自然环境中的微生物中快速克隆aroA 基因是非常高效和有利的。

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