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Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography

This protocol describes how to visualize the transient DNA compaction in cyanobacteria. DNA compaction is a dramatic cytoplasmic event recently found to occur in some cyanobacteria before cell division. However, due to the large cell size and the transient character, it is difficult to investigate t...

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Detalles Bibliográficos
Autores principales: Murata, Kazuyoshi, Kaneko, Yasuko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6126509/
https://www.ncbi.nlm.nih.gov/pubmed/30080205
http://dx.doi.org/10.3791/57197
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author Murata, Kazuyoshi
Kaneko, Yasuko
author_facet Murata, Kazuyoshi
Kaneko, Yasuko
author_sort Murata, Kazuyoshi
collection PubMed
description This protocol describes how to visualize the transient DNA compaction in cyanobacteria. DNA compaction is a dramatic cytoplasmic event recently found to occur in some cyanobacteria before cell division. However, due to the large cell size and the transient character, it is difficult to investigate the structure in detail. To overcome the difficulties, first, DNA compaction is reproducibly produced in the cyanobacterium Synechococcus elongatus PCC 7942 by synchronous culture using 12 h each light/dark cycle. Second, DNA compaction is monitored by fluorescence microscopy and captured by rapid freezing. Third, the detailed structure of DNA compacted cells is visualized in three dimensions (3D) by high voltage cryo-electron tomography. This set of methods is widely applicable to investigate transient structures in bacteria, e.g. cell division, chromosome segregation, phage infection etc., which are monitored by fluorescence microscopy and directly visualized by cryo-electron tomography at appropriate time points.
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spelling pubmed-61265092018-09-19 Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography Murata, Kazuyoshi Kaneko, Yasuko J Vis Exp Biology This protocol describes how to visualize the transient DNA compaction in cyanobacteria. DNA compaction is a dramatic cytoplasmic event recently found to occur in some cyanobacteria before cell division. However, due to the large cell size and the transient character, it is difficult to investigate the structure in detail. To overcome the difficulties, first, DNA compaction is reproducibly produced in the cyanobacterium Synechococcus elongatus PCC 7942 by synchronous culture using 12 h each light/dark cycle. Second, DNA compaction is monitored by fluorescence microscopy and captured by rapid freezing. Third, the detailed structure of DNA compacted cells is visualized in three dimensions (3D) by high voltage cryo-electron tomography. This set of methods is widely applicable to investigate transient structures in bacteria, e.g. cell division, chromosome segregation, phage infection etc., which are monitored by fluorescence microscopy and directly visualized by cryo-electron tomography at appropriate time points. MyJove Corporation 2018-07-17 /pmc/articles/PMC6126509/ /pubmed/30080205 http://dx.doi.org/10.3791/57197 Text en Copyright © 2018, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Biology
Murata, Kazuyoshi
Kaneko, Yasuko
Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography
title Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography
title_full Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography
title_fullStr Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography
title_full_unstemmed Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography
title_short Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography
title_sort visualization of dna compaction in cyanobacteria by high-voltage cryo-electron tomography
topic Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6126509/
https://www.ncbi.nlm.nih.gov/pubmed/30080205
http://dx.doi.org/10.3791/57197
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