Thursday, July 28, 2011: 8:30 AM
Bayside BC, 4th fl (Sheraton New Orleans)
Recent advances driven by research into the biology of model filamentous fungi such as Neurospora crassa and Aspergillus nidulans has generated gene targeting technologies that enable manipulation of all genes within many filamentous fungi of industrial importance. As a case study we have targeted the protein kinase, protein phosphatase, and nuclear pore protein encoding genes of A. nidulans for systematic deletion, protein purification by affinity purification and subcellular localization analysis. Advances in gene targeting and the availability of pre-made deletion constructs will be described making possible the global analysis of all A. nidulans genes. We will report unpublished deletion and characterization of all protein kinase, histidine kinase and PI3/PI4 kinase encoding genes, totaling 130 deletions, in this model fungus. Each gene has been deleted and defined as either essential or not essential. Non-essential haploid deleted strains have been tested for conditional phenotypes in response to numerous cellular stress conditions. In addition, for all 24 of the essential kinase genes, the terminal growth and cell cycle phenotype has been defined using heterokaryon rescue and microscopic analysis of DAPI stained cells. This global analysis has confirmed the phenotypes of previously studied kinase genes and has expanded the number of kinase genes that have been characterized in A. nidulans by 82. The deleted strains provide a powerful resource for analysis of all processes regulated by phosphorylation in fungi.