A comprehensive library of fluorescent transcriptional reporters for Escherichia coli.
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.
Zaslaver
Alon
A
Department of Molecular Cell Biology, Weizmann Institute of Science, 76100, Israel.
Bren
Anat
A
Ronen
Michal
M
Itzkovitz
Shalev
S
Kikoin
Ilya
I
Shavit
Seagull
S
Liebermeister
Wolfram
W
Surette
Michael G
MG
Alon
Uri
U
D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D019151:Peptide Library; D013050:Spectrometry, Fluorescence / Q000379:methods*; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics / Q000378:metabolism*
D029968:Escherichia coli Proteins; D019151:Peptide Library; D014157:Transcription Factors