Improved Cell-Free RNA and Protein Synthesis System

Li, Jun; Gu, Liangcai; Aach, John; Church, George M.

doi:10.1371/journal.pone.0106232

Improved Cell-Free RNA and Protein Synthesis System

Journal Article · Tue Sep 02 00:00:00 EDT 2014 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0106232· OSTI ID:1904697

Li, Jun ^[1]; Gu, Liangcai ^[2]; Aach, John ^[2]; Church, George M. ^[2]

Harvard Medical School, Boston, MA (United States); Harvard Medical School, Boston, MA (United States)
Harvard Medical School, Boston, MA (United States)

Cell-free RNA and protein synthesis (CFPS) is becoming increasingly used for protein production as yields increase and costs decrease. Advances in reconstituted CFPS systems such as the Protein synthesis Using Recombinant Elements (PURE) system offer new opportunities to tailor the reactions for specialized applications including in vitro protein evolution, protein microarrays, isotopic labeling, and incorporating unnatural amino acids. In this study, using firefly luciferase synthesis as a reporter system, we improved PURE system productivity up to 5 fold by adding or adjusting a variety of factors that affect transcription and translation, including Elongation factors (EF-Ts, EF-Tu, EF-G, and EF4), ribosome recycling factor (RRF), release factors (RF1, RF2, RF3), chaperones (GroEL/ES), BSA and tRNAs. The work provides a more efficient defined in vitro transcription and translation system and a deeper understanding of the factors that limit the whole system efficiency.

View Accepted Manuscript (DOE)

Research Organization:: Harvard Medical School, Boston, MA (United States); Harvard University, Cambridge, MA (United States)

Sponsoring Organization:: Merck KGaA; USDOE Office of Science (SC)

Grant/Contract Number:: FG02-02ER63445

OSTI ID:: 1904697

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 9 Vol. 9; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

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Improved Cell-Free RNA and Protein Synthesis System

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