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Harnessing single cell sorting to identify cell division genes and regulators in bacteria
Catherine Burke1, Michael Liu, Warwick Britton
1The ithree Institute, University of Technology, Sydney, New South Wales, Australia. Catherine.Burke@uts.edu.au
Plos One
|April 9, 2013
Summary
Researchers developed a new high-throughput screening method to identify bacterial cell division genes. This approach uses genomic libraries and flow cytometry to find novel regulators, aiding antibiotic development against pathogenic bacteria.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Cell division is a fundamental biological process crucial for organism growth and reproduction.
- Accurate spatial and temporal regulation of cell division relies on numerous proteins, some yet undiscovered.
- Identifying novel genes and regulators involved in bacterial cell division is vital for understanding fundamental biology and developing new antimicrobials.
Purpose of the Study:
- To develop and validate a novel, high-throughput screening method for identifying bacterial cell division genes and regulators.
- To discover previously unknown genes implicated in the cell division process.
- To establish a versatile screening strategy applicable to various bacterial species, including pathogens.
Main Methods:
- Construction and over-expression of a shotgun genomic expression library to perturb cell division in *Escherichia coli*.
- High-throughput screening using flow cytometry to isolate bacterial clones exhibiting altered cell division phenotypes (e.g., filamentous morphology).
- Genetic analysis of isolated clones to identify the responsible genes, including both known and novel cell division components.
Main Results:
- Successfully developed a novel high-throughput screening method for bacterial cell division genes.
- Identified bacterial clones with filamentous morphology, indicating defects in cell division.
- Discovered both known and previously unidentified genes involved in the cell division process in *Escherichia coli*.
- Demonstrated the applicability of the screening strategy to a model organism.
Conclusions:
- The developed high-throughput screening method is effective for identifying novel bacterial cell division genes and regulators.
- This approach significantly advances the discovery of genes involved in bacterial cell division.
- The method holds promise for identifying drug targets in pathogenic bacteria for novel antibiotic development.
