Improved Cell-Free RNA and Protein Synthesis System
View/ Open
Published Version
https://doi.org/10.1371/journal.pone.0106232Metadata
Show full item recordCitation
Li, Jun, Liangcai Gu, John Aach, and George M. Church. 2014. “Improved Cell-Free RNA and Protein Synthesis System.” PLoS ONE 9 (9): e106232. doi:10.1371/journal.pone.0106232. http://dx.doi.org/10.1371/journal.pone.0106232.Abstract
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.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4152126/pdf/Terms of Use
This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAACitable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:12987407
Collections
- HMS Scholarly Articles [17922]
Contact administrator regarding this item (to report mistakes or request changes)