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Methods Mol Biol. 2019;2004:223-238. doi: 10.1007/978-1-4939-9520-2_17.

Tracking Bacterial Chromosome Dynamics with Microfluidics-Based Live Cell Imaging.

Author information

1
National Centre for Biological Sciences, Tata Institute of Fundamental Research (TIFR), Bangalore, India.
2
Transdisciplinary University (TDU), Bangalore, India.
3
National Centre for Biological Sciences, Tata Institute of Fundamental Research (TIFR), Bangalore, India. anjana@ncbs.res.in.

Abstract

In bacteria, chromosomes are highly organized within the limited volume of the cell to form a nucleoid. Recent application of microscopy and chromosome conformation capture techniques have together provided a comprehensive understanding of the nature of this organization and the role of factors such as the structural maintenance of chromosomes (SMC) proteins in the establishment and maintenance of the same. In this chapter, we outline a microfluidics-based approach for live cell imaging of Escherichia coli chromosome dynamics in wild-type cells. This assay can be used to track the activity of the SMC complex, MukBEF, on DNA and assess the impact of perturbations such as DNA damage on chromosome organization and segregation.

KEYWORDS:

Chromosome; DNA damage; Escherichia coli; Fluorescence microscopy; Microfluidics; MukBEF; SMC proteins

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