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  Reconstitution and coupling of DNA replication and segregation in a biomimetic system

Hürtgen, D., Mascarenhas, J., Heymann, M., Murray, S., Schwille, P., & Sourjik, V. (2019). Reconstitution and coupling of DNA replication and segregation in a biomimetic system. ChemBioChem: A European Journal of Chemical Biology, 20(20), 2633-2642. doi:10.1002/cbic.201900299.

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© 2019 The Authors.

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 Creators:
Hürtgen, Daniel, Author
Mascarenhas, Judita, Author
Heymann, Michael1, Author              
Murray, Sean, Author
Schwille, Petra1, Author              
Sourjik, Victor, Author
Affiliations:
1Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              

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Free keywords: DNA nanoparticles DNA replication DNA segregation minimal cell ParM T7
 Abstract: A biomimetic system capable of replication and segregation of genetic material constitutes an essential component for the future design of a minimal synthetic cell. Here we have used the simple T7 bacteriophage system and the plasmid‐derived ParMRC system to establish in vitro DNA replication and DNA segregation, respectively. These processes were incorporated into biomimetic compartments providing an enclosed reaction space. The functional lifetime of the encapsulated segregation system could be prolonged by equipping it with ATP‐regenerating and oxygen‐scavenging systems. Finally, we showed that DNA replication and segregation processes could be coupled in vitro by using condensed DNA nanoparticles resulting from DNA replication. ParM spindles extended over tens of micrometers and could thus be used for segregation in compartments that are significantly longer than bacterial cell size. Overall, this work demonstrates the successful bottom‐up assembly and coupling of molecular machines that mediate replication and segregation, thus providing an important step towards the development of a fully functional minimal cell.

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Language(s): eng - English
 Dates: 20192019
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1002/cbic.201900299
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Title: ChemBioChem: A European Journal of Chemical Biology
  Other : ChemBioChem
Source Genre: Journal
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Publ. Info: Weinheim, Germany : Wiley-VCH
Pages: - Volume / Issue: 20 (20) Sequence Number: - Start / End Page: 2633 - 2642 Identifier: ISSN: 1439-4227
CoNE: https://pure.mpg.de/cone/journals/resource/110978984568897_1