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高通量方法在单细胞水平上鉴定细菌代谢产物产生菌的基因组变异。

A high-throughput approach to identify genomic variants of bacterial metabolite producers at the single-cell level.

机构信息

Institut für Bio- und Geowissenschaften, IBG-1: Biotechnologie, Forschungszentrum Jülich GmbH, D-52425 Jülich, Germany.

出版信息

Genome Biol. 2012 May 28;13(5):R40. doi: 10.1186/gb-2012-13-5-r40.

Abstract

We present a novel method for visualizing intracellular metabolite concentrations within single cells of Escherichia coli and Corynebacterium glutamicum that expedites the screening process of producers. It is based on transcription factors and we used it to isolate new L-lysine producing mutants of C. glutamicum from a large library of mutagenized cells using fluorescence-activated cell sorting (FACS). This high-throughput method fills the gap between existing high-throughput methods for mutant generation and genome analysis. The technology has diverse applications in the analysis of producer populations and screening of mutant libraries that carry mutations in plasmids or genomes.

摘要

我们提出了一种新的方法,可以可视化大肠杆菌和谷氨酸棒杆菌单个细胞内的代谢物浓度,从而加速了生产者的筛选过程。它基于转录因子,我们使用它通过荧光激活细胞分选 (FACS) 从大量诱变细胞文库中分离出谷氨酸棒杆菌的新型 L-赖氨酸生产突变体。这种高通量方法填补了现有高通量突变体生成和基因组分析方法之间的空白。该技术在分析生产者群体和筛选携带质粒或基因组突变的突变体文库方面具有多种应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b1c8/3446293/18d73be7ba41/gb-2012-13-5-r40-1.jpg

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