ArticleLife (Basel, Switzerland)2021
A Relationship between NTP and Cell Extract Concentration for Cell-Free Protein Expression.
Article in Life (Basel, Switzerland), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- A Cost-EffectiveBioengineering (Basel, Switzerland) · 2024Article
- Designing of an extract production protocol for industrial application of cell-free protein synthesis technology: Building from a current best practice to a quality by design approach.Engineering biology · 2023Review
- IRES-mediated Pichia pastoris cell-free protein synthesis.Bioresources and bioprocessing · 2023Article
- Rewiring cell-free metabolic flux inSynthetic biology (Oxford, England) · 2023Article
- What remains from living cells in bacterial lysate-based cell-free systems.Computational and structural biotechnology journal · 2023Review
- Trends and Outlooks in Synthetic Biology: A Special Issue for Celebrating 10 Years ofLife (Basel, Switzerland) · 2022Article
- System concentration shift as a regulator of transcription-translation system within liposomes.iScience · 2021Article
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Authors and funding
4 authors.
Funding
Abstract
The cell-free protein synthesis (CFPS) that synthesizes mRNA and protein from a template DNA has been featured as an important tool to emulate living systems in vitro. However, an obstacle to emulate living cells by CFPS is the loss of activity in the case of usage of high concentration cell extracts. In this study, we found that a high concentration of NTP which inhibits in the case of lower concentration cell extract restored the loss of CFPS activity using high concentration cell extracts. The NTP restoration was independent of the energy regeneration system used, and NTP derivatives also restored the levels of CFPS using a high concentration cell extract. Experiments using dialysis mode of CFPS showed that continuous exchange of small molecule reduced levels of NTP requirement and improved reaction speed of CFPS using the high concentration of cell extract. These findings contribute to the development of a method to understand the condition of living cells by in vitro emulation, and are expected to lead to the achievement of the reconstitution of living cells from biomolecule mixtures.
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Registered trials
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