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Visualization of DNA Compaction in Cyanobacteria by High-voltage Cryo-electron Tomography
- Source :
- Journal of Visualized Experiments.
- Publication Year :
- 2018
- Publisher :
- MyJove Corporation, 2018.
-
Abstract
- 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.
- Subjects :
- Cyanobacteria
Electron Microscope Tomography
General Immunology and Microbiology
biology
Cell division
General Chemical Engineering
General Neuroscience
DNA
macromolecular substances
biology.organism_classification
General Biochemistry, Genetics and Molecular Biology
Synchronous culture
chemistry.chemical_compound
chemistry
Cytoplasm
Biophysics
Fluorescence microscope
Humans
Cryo-electron tomography
Tomography
Subjects
Details
- ISSN :
- 1940087X
- Database :
- OpenAIRE
- Journal :
- Journal of Visualized Experiments
- Accession number :
- edsair.doi.dedup.....8d4608b20c9ca14c888e329d50c98051