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Probing gene expression in live cells, one protein molecule at a time.

Science (New York, N.Y.) | Mar 17, 2006

http://www.ncbi.nlm.nih.gov/pubmed/16543458

We directly observed real-time production of single protein molecules in individual Escherichia coli cells. A fusion protein of a fast-maturing yellow fluorescent protein (YFP) and a membrane-targeting peptide was expressed under a repressed condition. The membrane-localized YFP can be detected with single-molecule sensitivity. We found that the protein molecules are produced in bursts, with each burst originating from a stochastically transcribed single messenger RNA molecule, and that protein copy numbers in the bursts follow a geometric distribution. The quantitative study of low-level gene expression demonstrates the potential of single-molecule experiments in elucidating the workings of fundamental biological processes in living cells.

Pubmed ID: 16543458 RIS Download

Mesh terms: Bacterial Proteins | Blotting, Western | Escherichia coli K12 | Escherichia coli Proteins | Fluorescence | Gene Expression | Gene Expression Profiling | Luminescent Proteins | Membrane Proteins | Microscopy, Fluorescence | Molecular Probe Techniques | Poisson Distribution | Promoter Regions, Genetic | RNA, Bacterial | RNA, Messenger | Recombinant Fusion Proteins | Reverse Transcriptase Polymerase Chain Reaction | Stochastic Processes

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