Nature Methods 3, 623 - 628 (2006)
Published online: 21 July 2006; | doi:10.1038/nmeth895
A comprehensive library of fluorescent transcriptional reporters for Escherichia coliAlon Zaslaver1, Anat Bren1, Michal Ronen1, Shalev Itzkovitz1, Ilya Kikoin1, Seagull Shavit1, Wolfram Liebermeister2, Michael G Surette3 & Uri Alon11
Department of Molecular Cell Biology, Weizmann Institute of Science, 76100, Israel. 2
Berlin Center for Genome Based Bioinformatics, Max Planck Institute for Molecular Genetics, Ihnestr. 73, 14195 Berlin, Germany. 3
Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, AB, Canada T2N 4N1.
Correspondence should be addressed to Uri Alon urialon@weizmann.ac.il
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.
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