Date of Award

2013

Degree Type

Thesis

Department

Biological, Geological and Environmental Sciences

First Advisor

Boerner, G. Valentin

Subject Headings

Yeast, Meiosis, Saccharomyces cerevisiae, Budding yeast meiosis screen ZMM temperature-sensitive phenotype

Abstract

Budding yeast Saccharomyces cerevisiae contains a group of proteins named ZMM that constitutes a link between recombination and Synaptonemal Complex (SC) assembly. Yeast mutants that lack ZMM proteins have defects in recombination, SC formation and nuclear division progression. Meiotic cell cycle progression in zmm mutants is modulated by temperature. This conditional behavior is different at high and low temperatures. In my work so far, I have tried to identify new zmm-like genes involved in meiosis. To that end, I have carried out a genome-wide screen in the budding yeast S. cerevisiae. I have identified sporulation temperature sensitive zmm-like truncation mutants by using mini-transposon mediated random insertional mutagenesis approach. To confirm that the observed sporulation-temperature sensitive phenotype is caused by the transposon, a genetic outcross assay was carried out, and to determine the exact position of transposon integration in the yeast genome, direct genome sequencing was performed, followed by Southern hybridization. The defects that can potentially be detected by this genome wide screen approach are growth defects, defects in meiotic divisions and spore viability defect. Different classes of mutants have been identified, suggesting that insertional mutagenesis mediated genome wide screen is an appropriate genetic approach for identifying new genes involved in meiosis

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