Single-molecule fluorescence imaging of bacterial chromosome segregation dynamics in cell-free systems

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Cell division is a process of fundamental importance to all forms of life. During cell division, DNA must be properly segregated into each daughter cell in order to ensure the stable inheritance of genetic material. The aim of this project is to gain insight into the molecular mechanisms underpinning bacterial chromosome segregation from Vibrio cholerae as a target for novel antibiotics. The process of chromosome segregation in bacteria is regulated by Par (partition) proteins. Recent advances in fluorescence microscopy have revealed that bacterial chromosomes are highly organized and segregate with distinctive patterns in the cell. In this project, model systems consisting of nanopatterned arrays of biomolecules will be constructed and used as biomimetic platforms. These platforms will be utilized for the study of dynamics of partition proteins from Vibrio cholerae in order to better understand how they are involved in chromosome segregation. This project combines multidisciplinary approaches including single-molecule fluorescence microscopy, biochemistry, and bionanotechnology to visualize the purified proteins on synthetic surfaces in a cell-free system.