Measurement and Numerical Modeling of Cell-Free Protein Synthesis: Combinatorial Block-Variants of the PURE System

Paolo Carrara, Emiliano Altamura, Francesca D’Angelo, Fabio Mavelli and Pasquale Stano
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Paolo Carrara: Sciences Department, Roma Tre University, V.le G. Marconi 446, I-00146 Rome, Italy
Emiliano Altamura: Chemistry Department, University of Bari “Aldo Moro”, Via Orabona 4, I-70125 Bari, Italy
Francesca D’Angelo: Sciences Department, Roma Tre University, V.le G. Marconi 446, I-00146 Rome, Italy
Fabio Mavelli: Chemistry Department, University of Bari “Aldo Moro”, Via Orabona 4, I-70125 Bari, Italy
Pasquale Stano: Sciences Department, Roma Tre University, V.le G. Marconi 446, I-00146 Rome, Italy

Data, 2018, vol. 3, issue 4, 1-12

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).

Keywords: synthetic cells; synthetic biology; in vitro protein synthesis; transcription–translation (TX–TL); extrinsic stochastic effects; solute partition; PURE system (search for similar items in EconPapers)
JEL-codes: C8 C80 C81 C82 C83 (search for similar items in EconPapers)
Date: 2018
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