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Related Experiment Video

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Live Cell Imaging of Chromosome Segregation During Mitosis
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Tracking Bacterial Chromosome Dynamics with Microfluidics-Based Live Cell Imaging.

Suchitha Raghunathan1,2, Anjana Badrinarayanan3

  • 1National Centre for Biological Sciences, Tata Institute of Fundamental Research (TIFR), Bangalore, India.

Methods in Molecular Biology (Clifton, N.J.)
|June 1, 2019
PubMed
Summary

This study presents a microfluidics method to observe bacterial chromosome organization and the function of structural maintenance of chromosomes (SMC) proteins in live Escherichia coli cells. The assay tracks SMC complex activity and DNA damage effects on chromosome dynamics.

Keywords:
ChromosomeDNA damageEscherichia coliFluorescence microscopyMicrofluidicsMukBEFSMC proteins

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Cell Biology

Background:

  • Bacterial chromosomes form a highly organized nucleoid within the cell.
  • Structural Maintenance of Chromosomes (SMC) proteins are crucial for chromosome organization and dynamics.
  • Understanding chromosome organization is key to bacterial cell viability.

Purpose of the Study:

  • To outline a microfluidics-based approach for live cell imaging of bacterial chromosome dynamics.
  • To track the activity of the SMC complex, MukBEF, on DNA in Escherichia coli.
  • To assess the impact of DNA damage on chromosome organization and segregation.

Main Methods:

  • Live cell imaging using a microfluidics-based assay.
  • Observation of wild-type Escherichia coli cells.
  • Tracking of the MukBEF SMC complex activity on DNA.

Main Results:

  • The developed microfluidics assay enables visualization of bacterial chromosome dynamics in real-time.
  • The assay allows for the assessment of SMC complex (MukBEF) activity on DNA.
  • The system can evaluate the effects of perturbations like DNA damage on chromosome organization.

Conclusions:

  • This microfluidics approach provides a powerful tool for studying bacterial chromosome organization and dynamics.
  • It facilitates the investigation of SMC protein function in response to cellular stress.
  • The method contributes to a deeper understanding of how bacteria maintain genome integrity.