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
;
2018-01-01
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).I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
