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Cited by 5RNA and protein synthesis mechanismsMicrobial Metabolic Engineering and BioproductionBacterial Genetics and Biotechnology

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 access
Meyerowitz, Joseph T., Larsson, Elin M., Murray, Richard M.·2024·ACS Synthetic Biology
DOI 10.1021/acssynbio.3c00468

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Abstract

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.

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Metadata

Authors
Meyerowitz, J. T., Larsson, E. M., & Murray, R. M.
Affiliations
California Institute of Technology (US)
Corresponding author
Meyerowitz, Joseph T.
ORCID
Venue
ACS Synthetic Biology
Publisher
American Chemical Society (ACS)
Published
2024-02-06
Volume
13
Issue
2
Pages
530-537
ISSN
2161-5063 (ISSN-L)
DOI
10.1021/acssynbio.3c00468
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
Licence
Locations
  • ACS Synthetic Biology (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Army Research Office (W911NF-09-0001)
  • Division of Chemical, Bioengineering, Environmental, and Transport Systems (CBET-1903477)
  • Institute for Collaborative Biotechnologies (W911NF-19-2-0026, W911NF-19-D-0001)
  • International Human Frontiers Science Program (International Human Frontiers Science Program)
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

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