Single Molecule Studies of Replisomal Function

The replisome is the complex molecular machine that faithfully makes copies of DNA molecules in cells that are about to divide. Bacterial replisomes that replicate DNA in vitro at the same rates as inside the cell can be assembled from their many separately-isolated protein subunits. New techniques allow study of replisome-mediated DNA replication at the single molecule level, to probe the activities of complete replisomes and functional subassemblies. Two complementary techniques will be applied, a flow-visualisation method in the USA, and a force-measuring optical tweezers technique in Australia. These will be used in ongoing work, extending from the simpler replisome of phage T7 to the complete bacterial system