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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...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2018
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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. |
format | Online Article Text |
id | pubmed-6126509 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
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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