My research is directed towards understanding the proteins and mechanisms involved in the recombination of homologous DNA molecules. Homologous recombination has an important role in facilitating cell survival following damage to DNA; defects in recombination repair mechanisms can lead to genomic instability and initiation of carcinogenesis.
The goal of my current research is to develop a complete in vitro recombination system with purified human proteins to facilitate a greater understanding of recombination mediated DNA repair in vivo.
We are expressing in E.coli human proteins implicated in homologous recombination and purifying them by column chromatography. Activities of proteins consistent with a role in homologous recombination are assayed using a variety of techniques. This includes electrophoretic assays for DNA binding and strand exchange, thin layer chromatography assays for ATPase activity, and visualisation of protein-DNA complexes by transmission electron microscopy. In addition we are raising antibodies to purified proteins to provide tools to further investigate the inter-relationships between different recombination proteins in cellular immunolocalisation studies and by co-immunoprecipitation.