Protein synthesis is at the core of bottom-up construction of artificial cellular mimics. Intriguingly, several reports have revealed that when a transcription-translation (TX-TL) kit is encapsulated inside lipid vesicles (or water-in-oil droplets), high between-vesicles diversity is observed in terms of protein synthesis rate and yield. Stochastic solute partition can be a major determinant of these observations. In order to verify that the variation of TX-TL components concentration brings about a variation of produced protein rate and yield, here we directly measure the performances of the PURE system' TX-TL kit variants. We report and share the kinetic traces of the enhanced Green Fluorescent Protein (eGFP) synthesis in bulk aqueous phase, for 27 combinatorial block-variants. The eGFP production is a sensitive function of TX-TL components concentration in the explored concentration range. Providing direct evidence that protein synthesis yield and rate actually mirror the TX-TL composition, this study supports the above-mentioned hypothesis on stochastic solute partition, without excluding, however, the contribution of other factors (e.g., inactivation of components).

Measurement and numerical modeling of cell-free protein synthesis: Combinatorial block-variants of the pure system

Altamura E.
Investigation
;
Mavelli F.
Writing – Review & Editing
;
2018

Abstract

Protein synthesis is at the core of bottom-up construction of artificial cellular mimics. Intriguingly, several reports have revealed that when a transcription-translation (TX-TL) kit is encapsulated inside lipid vesicles (or water-in-oil droplets), high between-vesicles diversity is observed in terms of protein synthesis rate and yield. Stochastic solute partition can be a major determinant of these observations. In order to verify that the variation of TX-TL components concentration brings about a variation of produced protein rate and yield, here we directly measure the performances of the PURE system' TX-TL kit variants. We report and share the kinetic traces of the enhanced Green Fluorescent Protein (eGFP) synthesis in bulk aqueous phase, for 27 combinatorial block-variants. The eGFP production is a sensitive function of TX-TL components concentration in the explored concentration range. Providing direct evidence that protein synthesis yield and rate actually mirror the TX-TL composition, this study supports the above-mentioned hypothesis on stochastic solute partition, without excluding, however, the contribution of other factors (e.g., inactivation of components).
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11586/250721
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