Kinetics of the cellular intake of a gene expression inducer at high concentrations

被引:14
作者
Tran, Huy [1 ]
Oliveira, Samuel M. D. [1 ]
Goncalves, Nadia [1 ]
Ribeiro, Andre S. [1 ]
机构
[1] Tampere Univ Technol, Dept Signal Proc, Lab Biosyst Dynam, FI-33101 Tampere, Finland
基金
芬兰科学院;
关键词
IN-VIVO KINETICS; ESCHERICHIA-COLI; SINGLE-CELL; TRANSCRIPTION INITIATION; SEQUENTIAL MECHANISM; RNA-POLYMERASE; LAC REPRESSOR; TIME; PROMOTERS; INDUCTION;
D O I
10.1039/c5mb00244c
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
From in vivo single-event measurements of the transient and steady-state transcription activity of a single-copy lac-ara-1 promoter in Escherichia coli, we characterize the intake kinetics of its inducer (IPTG) from the media. We show that the empirical data are well-fit by a model of intake assuming a bilayer membrane, with the passage through the second layer being rate-limiting, coupled to a stochastic, sub-Poissonian, multi-step transcription process. Using this model, we show that for a wide range of extracellular inducer levels (up to 1.25 mM) the intake process is diffusive-like, suggesting unsaturated membrane permeability. Inducer molecules travel from the periplasm to the cytoplasm in, on average, 31.7 minutes, strongly affecting cells' response time. The novel methodology followed here should aid the study of cellular intake mechanisms at the single-event level.
引用
收藏
页码:2579 / 2587
页数:9
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