Constructing Cell-Free Expression Systems for Low-Cost Access
Fernando Guzmán‐Chávez, Aníbal Arce, Abhinav Adhikari, Sandra Vadhin, José Antonio Pedroza‐García, Chiara Gandini, Jim W. Ajioka, Jenny Molloy, Sobeida Sánchez‐Nieto, Jeffrey D. Varner, Fernán Federici, Jim Haseloff
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.