Development of a System for the Isolation of Genes Involved in Cold Adaptation in Antarctic Yeasts

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The extreme environment of Antarctica provides a unique opportunity to study adaptation of organisms to cold as well as dry conditions. This research will focus on an Antarctic yeast (Leucosporidium sp.) as a model system to study cellular function and regulation at the molecular level through the isolation of genes required for cold adaptation. Before genes necessary to examine these processes will be cloned, a transformational system, including a mechanism for the recovery of isolated DNA, will be developed for Leucosporidium. Antibiotic resistance markers on plasmid or cosmid vectors will be utilized to follow the introduction of exogenous DNA into this system and plasmid or cosmid DNA will be re-isolated from the transformed yeast to insure replication fidelity after transformation. This project will lead to the application of the techniques of RNA subtraction and complementation of induced mutations to isolate those genes which are either constitutively regulated by life in the cold or variably regulated for adaptation to a changing cold environment.