The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
Development of cell-free transcription-translation systems in three soil Pseudomonads
Open accessPreprint — published as 10.1021/acssynbio.3c00468 · Paper page
Abstract
In vitro transcription-translation (TX-TL) can enable faster engineering of biological systems. This speed-up can be significant, especially in difficult-to-transform chassis. This work shows the successful development of TX-TL systems using three soil-derived wild-type Pseudomonads known to promote plant growth: Pseudomonas synxantha, Pseudomonas chlororaphis , and Pseudomonas aureofaciens . One, P. synxantha , was further characterized. A lysate test of P. synxantha showed a maximum protein yield of 2.5 μ M at 125 proteins per DNA template and a maximum protein synthesis rate of 20 nM/min. A set of different constitutive promoters driving mNeonGreen expression were tested in TX-TL and integrated into the genome, showing similar normalized strengths for in vivo and in vitro fluorescence. This correspondence between the TX-TL derived promoter strength and the in vivo promoter strength indicates these lysate-based cell-free systems can be used to characterize and engineer biological parts without genome integration, enabling a faster designbuild-test cycle.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764
Flagged first, then by citations
Research works printed but not linked: 45
- 2009, Enzymatic assembly of DNA molecules up to several hundred kilobases, Nature methods, 6, 343, 10.1038/nmeth.1318 · DOI
- 10.1038/nbt.3659 · DOI · Paper page
- 10.1038/nature04640 · DOI · Paper page
- 2022, A microbial supply chain for production of the anti-cancer drug vinblastine, Nature, 609, 341, 10.1038/s41586-022-05157-3 · DOI
- 2011, Engineering modular and orthogonal genetic logic gates for robust digital-like synthetic biology, Nature communications, 2, 508, 10.1038/ncomms1516 · DOI
- 10.15252/msb.20156663 · DOI
- 2020, Synthetic biology enables programmable cell-based biosensors, ChemPhysChem, 21, 132, 10.1002/cphc.201900739 · DOI
- 10.1101/cshperspect.a023812 · DOI · Paper page
- 10.1093/nar/gkab1135 · DOI
- 10.1126/science.aac7341 · DOI
- 2019, Escherichia coli “marionette” strains with 12 highly optimized small-molecule sensors, Nature Chemical Biology, 15, 196, 10.1038/s41589-018-0168-3 · DOI
- 10.1038/nbt1413 · DOI
- 10.1128/AEM.69.11.6703-6714.2003 · DOI
- 10.1093/femsre/fuv006 · DOI
- 2021, Deciphering bacterial mechanisms of root colonization, Environmental Microbiology Reports, 13, 428, 10.1111/1758-2229.12934 · DOI
- 10.1002/ppul.21011 · DOI
- 10.1038/s41579-021-00567-y · DOI
- 2020, Development of a genetic toolset for the highly engineerable and metabolically versatile Acine-tobacter baylyi ADP1, Nucleic Acids Research, 48, 5169, 10.1093/nar/gkaa167 · DOI
- 2020, Genetic circuit design automation for the gut resident species bacteroides thetaiotaomicron, Nature Biotechnology, 38, 962, 10.1038/s41587-020-0468-5 · DOI
- 10.1016/j.cell.2017.03.041 · DOI
- L. E. Leiva and A. Katz , “Regulation of leaderless mRNA translation in bacteria,” Microorganisms, vol. 10, Mar. 2022. · DOI
- 2020, Methodologies for preparation of prokaryotic extracts for cell-free expression systems, Synthetic and Systems Biotechnology, 5, 252, 10.1016/j.synbio.2020.07.006 · DOI
- 10.1073/pnas.1715806115 · DOI
- S. J. Moore , H.-E. Lai , J. Li , and P. S. Freemont , “Streptomyces cell-free systems for natural product discovery and engineering,” Natural product reports, 2023. · DOI
- 10.1128/jb.170.8.3499-3508.1988 · DOI
- 10.1038/nrmicro1129 · DOI
- 10.1126/science.abd1515 · DOI
- 10.1007/s00253-004-1642-2 · DOI
- 2013, Isolation and characterization of bacteria capable of metabolizing lignin-derived low molecular weight compounds, Biotechnology and bioprocess engineering, 18, 736, 10.1007/s12257-012-0807-6 · DOI
- 10.1002/jobm.3620370309 · DOI
- I. Mahdi , N. Fahsi , M. Hijri , and M. Sobeh , “Antibiotic resistance in plant growth promoting bacteria: A comprehensive review and future perspectives to mitigate potential gene invasion risks,” Frontiers in Microbiology, vol. 13, 2022. · DOI
- 2017, Enzymatic degradation of lignin in soil: a review, Sustainability, 9, 1163, 10.3390/su9071163 · DOI
- 10.1007/s11274-017-2356-9 · DOI
- 10.1038/nrmicro1129 · DOI
- K. Nesemann , S. A. Braus-Stromeyer , A. Thuermer , R. Daniel , D. V. Mavrodi , L. S. Thomashow , D. M. Weller , and G. H. Braus , “Draft genome sequence of the phenazine-producing Pseudomonas fluorescens strain 2-79,” Genome Announc., vol. 3, Mar. 2015. · DOI
- 10.1094/MPMI.2000.13.12.1293 · DOI
- 2019, TYGS is an automated high-throughput platform for state-of-the-art genome-based taxonomy, Nature Communications, 10, 2182, 10.1038/s41467-019-10210-3 · DOI
- 10.1038/nrmicro3253 · DOI
- Z. Z. Sun , C. A. Hayes , J. Shin , F. Caschera , R. M. Murray , and V. Noireaux , “Protocols for implementing an escherichia coli based TX-TL cell-free expression system for synthetic biology,” J. Vis. Exp., p. e50762, Sept. 2013. · DOI
- 2014, Linear DNA for rapid prototyping of synthetic biological circuits in an Escherichia coli based tx-tl cell-free system, ACS Synthetic Biology, 3, 387, 10.1021/sb400131a · DOI
- 10.1073/pnas.85.23.8973 · DOI
- 10.1128/AEM.02041-16 · DOI
- 2015, Tn7-based device for calibrated heterologous gene expression in Pseudomonas putida, ACS Synthetic Biology, 4, 1341, 10.1021/acssynbio.5b00058 · DOI
- 2006, mini-tn 7 insertion in bacteria with single att tn 7 sites: example Pseudomonas aeruginosa, Nature Protocols, 1, 153, 10.1038/nprot.2006.24 · DOI
- 2005, A tn 7-based broad-range bacterial cloning and expression system, Nature Methods, 2, 443, 10.1038/nmeth765 · DOI
Metadata
- Authors
- Meyerowitz, J. T., Larsson, E. M., & Murray, R. M.
- Affiliations
- California Institute of Technology (US)
- Corresponding author
- Meyerowitz, Joseph T.
- Venue
- ACS Synthetic Biology
- Publisher
- openRxiv
- Published
- 2023-06-09
- Volume
- 13
- Issue
- 2
- Pages
- 530-537
- ISSN
- 2161-5063 (ISSN-L)
- OpenAlex
- W4391565761
- Cited by
- 5
- References
- 1
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 1
- FWCI
- 0.68
- Type
- journal article
- Language
- en
- Open access
- green
- dx.doi.org
- Licence
- cc-by
- Locations
- ACS Synthetic Biology (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Science Foundation (1903477, W911NF-19)
- Division of Chemical, Bioengineering, Environmental, and Transport Systems (CBET-1903477)
- Army Research Office (W911NF-09-0001, W911NF-19-2-0026, W911NF‐19‐D‐0001, W911NF, W911NF-19)
- Institute for Collaborative Biotechnologies (W911NF‐09‐0001, W911NF-19-D-0001, W911NF‐19‐2‐0026)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- cell-free transcription-translation, cell-free protein synthesis, Pseudomonas synxantha, Pseudomonas chlororaphis, Pseudomonas aureofaciens, plant growth-promoting bacteria, design-build-test cycle, chassis engineering, protein yield
- MeSH terms
- Cell-Free System, DNA, Escherichia coli, Transcription, Genetic, Protein Biosynthesis
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
