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Appl Environ Microbiol. 2011 May;77(9):2863-8. doi: 10.1128/AEM.01915-10. Epub 2011 Mar 4.

LacI(Ts)-regulated expression as an in situ intracellular biomolecular thermometer.

Author information

1
Department of Civil, Environmental, and Architectural Engineering, University of Colorado at Boulder, 1111 Engineering Dr., UCB Box 428, Boulder, CO 80309, USA.

Abstract

In response to needs for in situ thermometry, a temperature-sensitive vector was adapted to report changes in the intracellular heat content of Escherichia coli in near-real time. This model system utilized vectors expressing increasing quantities of β-galactosidase in response to stepwise temperature increases through a biologically relevant range (22 to 45°C). As judged by calibrated fluorometric and colorimetric reporters, both whole E. coli cells and lysates expressed significant repeatable changes in β-galactosidase activity that were sensitive to temperature changes of less than 1°C (35 to 45°C). This model system suggests that changes in cellular heat content can be detected independently of the medium in which cells are maintained, a feature of particular importance where the medium is heterogeneous or nonaqueous, or otherwise has a low heat transfer capacity. We report here that the intracellular temperature can be reliably obtained in near-real time using reliable fluorescent reporting systems from cellular scales, with a 20°C range of detection and at least 0.7°C sensitivity between 35 and 45°C.

PMID:
21378059
PMCID:
PMC3126416
DOI:
10.1128/AEM.01915-10
[Indexed for MEDLINE]
Free PMC Article

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