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Science 2 January 1998:
Vol. 279. no. 5347, pp. 84 - 88
DOI: 10.1126/science.279.5347.84
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Reports

Quantitation of Transcription and Clonal Selection of Single Living Cells with beta -Lactamase as Reporter

Gregor Zlokarnik, Paul A. Negulescu, Thomas E. Knapp, Lora Mere, Neal Burres, * Luxin Feng, Michael Whitney, Klaus Roemer, Roger Y. Tsien dagger

Gene expression was visualized in single living mammalian cells with beta -lactamase as a reporter that hydrolyzes a substrate loaded intracellularly as a membrane-permeant ester. Each enzyme molecule changed the fluorescence of many substrate molecules from green to blue by disrupting resonance energy transfer. This wavelength shift was detectable by eye or color film in individual cells containing less than 100 beta -lactamase molecules. The robust change in emission ratio reveals quantitative heterogeneity in real-time gene expression, enables clonal selection by flow cytometry, and forms a basis for high-throughput screening of pharmaceutical candidate drugs in living mammalian cells.

G. Zlokarnik, P. A. Negulescu, T. E. Knapp, L. Mere, N. Burres, L. Feng, M. Whitney, Aurora Biosciences, 11010 Torreyana Road, San Diego, CA 92121, USA.
K. Roemer, University of the Saarland, D-66421 Homburg/Saar, Germany.
R. Y. Tsien, Howard Hughes Medical Institute and Department of Pharmacology, University of California at San Diego, La Jolla, CA 92093-0647, USA.
*   Present address: 11757 Spruce Run Drive, San Diego, CA 92131-3709, USA.

dagger    To whom correspondence should be addressed.

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