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Issue 5, 2012
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Experiment and mathematical modeling of gene expression dynamics in a cell-free system

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Abstract

Cell-free in vitro expression is increasingly important for high-throughput expression screening, high yield protein production and synthetic biology applications. Yet its potential for quantitative investigation of gene expression and regulatory circuits is limited by the availability of data on composition, kinetic rate constants and standardized computational tools for modeling. Here we report on calibration measurements and mathematical modeling of a reconstituted in vitro expression system. We measured a series of GFP expression and mRNA transcription time courses under various initial conditions and established the translation step as the bottle neck of in vitro protein synthesis. Cell-free translation was observed to expire after 3 h independent of initial template DNA concentration. We developed a minimalistic rate equation model and optimized its parameters by performing a concurrent fit to measured time courses. The model predicts the dependence of protein yield not only on template DNA concentration, but also on experimental timing and hence is a valuable tool to optimize yield strategies.

Graphical abstract: Experiment and mathematical modeling of gene expression dynamics in a cell-free system

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Publication details

The article was received on 08 Sep 2011, accepted on 14 Mar 2012 and first published on 05 Apr 2012


Article type: Paper
DOI: 10.1039/C2IB00102K
Citation: Integr. Biol., 2012,4, 494-501
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    Experiment and mathematical modeling of gene expression dynamics in a cell-free system

    T. Stögbauer, L. Windhager, R. Zimmer and J. O. Rädler, Integr. Biol., 2012, 4, 494
    DOI: 10.1039/C2IB00102K

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