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Sökning: WFRF:(Olieric Natacha)

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1.
  • Bieniossek, Christoph, et al. (författare)
  • Automated unrestricted multigene recombineering for multiprotein complex production
  • 2009
  • Ingår i: Nature Methods. - 1548-7091 .- 1548-7105. ; 6:6, s. 447-U68
  • Tidskriftsartikel (refereegranskat)abstract
    • Structural and functional studies of many multiprotein complexes depend on recombinant-protein overexpression. Rapid revision of expression experiments and diversification of the complexes are often crucial for success of these projects; therefore, automation is increasingly indispensable. We introduce Acembl, a versatile and automatable system for protein-complex expression in Escherichia coli that uses recombineering to facilitate multigene assembly and diversification. We demonstrated protein-complex expression using Acembl, including production of the complete prokaryotic holotranslocon.
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2.
  • Martiel, Isabelle, et al. (författare)
  • Versatile microporous polymer-based supports for serial macromolecular crystallography
  • 2021
  • Ingår i: Acta Crystallographica Section D. - 2059-7983. ; 77, s. 1153-1167
  • Tidskriftsartikel (refereegranskat)abstract
    • Serial data collection has emerged as a major tool for data collection at state-of-the-art light sources, such as microfocus beamlines at synchrotrons and X-ray free-electron lasers. Challenging targets, characterized by small crystal sizes, weak diffraction and stringent dose limits, benefit most from these methods. Here, the use of a thin support made of a polymer-based membrane for performing serial data collection or screening experiments is demonstrated. It is shown that these supports are suitable for a wide range of protein crystals suspended in liquids. The supports have also proved to be applicable to challenging cases such as membrane proteins growing in the sponge phase. The sample-deposition method is simple and robust, as well as flexible and adaptable to a variety of cases. It results in an optimally thin specimen providing low background while maintaining minute amounts of mother liquor around the crystals. The 2 × 2 mm area enables the deposition of up to several microlitres of liquid. Imaging and visualization of the crystals are straightforward on the highly transparent membrane. Thanks to their affordable fabrication, these supports have the potential to become an attractive option for serial experiments at synchrotrons and free-electron lasers.
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