A comprehensive library of fluorescent transcriptional reporters for Escherichia coli
A comprehensive library of fluorescent transcriptional reporters for Escherichia coli
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DOI：
10.1038/nmeth895
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发表时间：
2006-08-01
期刊：
影响因子：
48
通讯作者：
Alon, Uri
中科院分区：
文献类型：
--
作者：
Zaslaver, Alon;Bren, Anat;Alon, Uri
大肠杆菌被广泛用于系统生物学研究；然而，需要有能够用于准确和全面地测量单个活细胞中表达动态的工具。为了解决这个问题，我们构建了一个在大肠杆菌K12中约2000个不同启动子各自与绿色荧光蛋白（gfp）转录融合的文库，涵盖了该生物体中绝大多数启动子。每个启动子融合体由一个低拷贝质粒表达。我们证明，在一个葡萄糖 - 乳糖二次生长转换实验中，这个文库可用于在基因组规模上获得对启动子活性的高精度动态测量。该文库能够检测到大肠杆菌中约80个以前未被鉴定的转录单元，包括在以前已知的操纵子（如乳糖操纵子）内假定的内部启动子。这个文库可作为一种工具，用于对活的大肠杆菌细胞中的转录网络进行准确、高分辨率的分析。
E. coli is widely used for systems biology research; there exists a need, however, for tools that can be used to accurately and comprehensively measure expression dynamics in individual living cells. To address this we present a library of transcriptional fusions of gfp to each of about 2,000 different promoters in E. coli K12, covering the great majority of the promoters in the organism. Each promoter fusion is expressed from a low-copy plasmid. We demonstrate that this library can be used to obtain highly accurate dynamic measurements of promoter activity on a genomic scale, in a glucose-lactose diauxic shift experiment. The library allowed detection of about 80 previously uncharacterized transcription units in E. coli, including putative internal promoters within previously known operons, such as the lac operon. This library can serve as a tool for accurate, high-resolution analysis of transcription networks in living E. coli cells.