Improved cell-free RNA and protein synthesis system.
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 spe...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4152126?pdf=render |
| _version_ | 1828342992017031168 |
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| author | Jun Li Liangcai Gu John Aach George M Church |
| author_facet | Jun Li Liangcai Gu John Aach George M Church |
| author_sort | Jun Li |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T23:36:53Z |
| format | Article |
| id | doaj.art-33fd31997b8446fa91cca38afefbf3c6 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T23:36:53Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-33fd31997b8446fa91cca38afefbf3c62022-12-22T02:24:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0199e10623210.1371/journal.pone.0106232Improved cell-free RNA and protein synthesis system.Jun LiLiangcai GuJohn AachGeorge M ChurchCell-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.http://europepmc.org/articles/PMC4152126?pdf=render |
| spellingShingle | Jun Li Liangcai Gu John Aach George M Church Improved cell-free RNA and protein synthesis system. PLoS ONE |
| title | Improved cell-free RNA and protein synthesis system. |
| title_full | Improved cell-free RNA and protein synthesis system. |
| title_fullStr | Improved cell-free RNA and protein synthesis system. |
| title_full_unstemmed | Improved cell-free RNA and protein synthesis system. |
| title_short | Improved cell-free RNA and protein synthesis system. |
| title_sort | improved cell free rna and protein synthesis system |
| url | http://europepmc.org/articles/PMC4152126?pdf=render |
| work_keys_str_mv | AT junli improvedcellfreernaandproteinsynthesissystem AT liangcaigu improvedcellfreernaandproteinsynthesissystem AT johnaach improvedcellfreernaandproteinsynthesissystem AT georgemchurch improvedcellfreernaandproteinsynthesissystem |