Constructing Cell-Free Expression Systems for Low-Cost Access
Fernando Guzmán‐Chávez(University of Cambridge), Aníbal Arce(Pontificia Universidad Católica de Chile), Abhinav Adhikari(Cornell University), Sandra Vadhin(Cornell University), José Antonio Pedroza‐García(Universidad Nacional Autónoma de México), Chiara Gandini(University of Cambridge), Jim W. Ajioka(University of Cambridge), Jenny Molloy(University of Cambridge), Sobeida Sánchez‐Nieto(Universidad Nacional Autónoma de México), Jeffrey D. Varner(Cornell University), Fernán Federici(Pontificia Universidad Católica de Chile), Jim Haseloff(University of Cambridge)
Cited by 61Open Access
Abstract
I. Third, we demonstrated the endogenous regeneration of nucleoside triphosphates and synthesis of pyruvate in cell-free systems (CFSs) based on phosphoenol pyruvate (PEP) and maltodextrin (MDX). We exploited this novel finding to demonstrate the use of a cell-free mixture completely free of any exogenous nucleotide triphosphates (NTPs) to generate high yields of sfGFP expression. Together, these modifications can produce desiccated extracts that are 203-424-fold cheaper than commercial versions. These improvements will facilitate wider use of CFS for research and education purposes.
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