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Identification and analysis of genes involved in kinetochore function during mitosis in caenorhabditis elegans


Research objectives and content Although the mechanistic and cell cycle signalling activities of higher eukaryotic kinetochores are now well-documented, the underlying molecules and mechanisms remain poorly understood. The goal of the proposed research is to use C. elegans genetic, molecular and cytological analyses to better understand the mechanisms of chromosome segregation at mitosis. First, I will identify and characterize new genes involved in kinetochore function during mitosis. Complementing Dr. Hyman's ongoing screen for spindle function mutants, I will specifically identify kinetochore function mutants using time-lapse fluorescence microscopy to simultaneously image GFP-tagged chromosomes and rhodamine-tubulin-marked microtubules in living cells. The identified genes will be cloned and further analyzed. Second, I will also study C. elegans homologues of known vertebrate kinetochore genes using existing EST clones to generate antibodies for immunocytochemical studies and initiate an antisense-based analysis of their functions in vivo. Training content (objective, benefit and expected impact) Although I have strong training in the study of the cytoskeleton and mitosis in vertebrate cells using molecular, biochemical and immunocytochemical methods, I have little experience using genetic analyses of whole organisms, and have never studied C. elegans. The proposed project will therefore integrate thematic continuity with new technical and theoretical training. I expect this to greatly benefit me by imparting the necessary tools to initiate my own independent research program within a few years.
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