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Cell cycle synchronisation of Trypanosoma brucei by centrifugal counter-flow elutriation reveals the timing of nuclear and kinetoplast DNA replication

Benz, Corinna and Dondelinger, Frank and McKean, Paul Gerard and Urbaniak, Michael Daniel (2017) Cell cycle synchronisation of Trypanosoma brucei by centrifugal counter-flow elutriation reveals the timing of nuclear and kinetoplast DNA replication. Scientific Reports, 7. ISSN 2045-2322

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    Abstract

    We report an optimised centrifugal counter-flow elutriation protocol for the rapid and direct isolation of G1 cell cycle synchronised populations of both the procyclic and bloodstream form stages of Trypanosoma brucei that yields viable and proliferative cells. The high quality of the synchronisation achieved can be judged by the uniform DNA content, narrow size distribution, synchronous division, and the maintenance of synchronicity into subsequent cell cycles. We show that early-eluting fractions represent different G1 subpopulations that progress through the cell cycle with distinct temporal profiles post-elutriation, as exemplified by the observation of the maturation of a second flagellar basal body in late G1 phase, DNA replication in S phase, and dimethylation of histone H3 in mitosis/cytokinesis. We use our temporal observations to construct a revised model of the relative timing and duration of the nuclear and kinetoplast cell cycle that differs from the current model.

    Item Type: Journal Article
    Journal or Publication Title: Scientific Reports
    Uncontrolled Keywords: /dk/atira/pure/subjectarea/asjc/1000
    Subjects: ?? DNA REPLICATIONISOLATION, SEPARATION AND PURIFICATIONPARASITE BIOLOGYGENERAL ??
    Departments: Faculty of Health and Medicine > Biomedical & Life Sciences
    Faculty of Health and Medicine > Medicine
    ID Code: 88798
    Deposited By: ep_importer_pure
    Deposited On: 20 Nov 2017 12:48
    Refereed?: Yes
    Published?: Published
    Last Modified: 26 May 2019 02:12
    Identification Number:
    URI: http://eprints.lancs.ac.uk/id/eprint/88798

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