Use of fluorescently tagged SSB proteins in in vivo localization experiments.

The time and place of DNA replication in cells provides invaluable insight into the cell cycle and DNA metabolism. An effective means of obtaining this information is through fluorescence microscopy. The abundance of Single-Strand Binding protein, SSB, at the replication fork makes it a good reporter of DNA replication. In this chapter I describe how to observe replication of the Escherichia coli chromosome in a strain that synthesizes a fluorescent derivative of SSB. This methodology provides information about the position and dynamics of DNA replication through epifluorescence.