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Once a set of mutations affecting one pathway or

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Unformatted text preview: ). Once a set of mutations affecting one pathway or process has been identified in the haploid products, mutants are crossed to produce a diploid cell for complementation testing. The diploid containing two different mutations is then sporulated and during meiosis the chromosomes are reshuffled by independent assortment and recombination. Some of the haploid spores will contain both mutations. If the mutations are in different genes, the cells will have the wild-type phenotype. (See Chapter 5 of the main textbook for more on genetic analysis in yeast using tetrad analysis.) The ability to easily isolate many mutants and the development of clever mutant screens makes possible the sophisticated analysis of gene function and the interaction of several genes in a pathway. Dominant gain-of-function mutations are particularly useful for pathway analysis using epistasis tests. Consider a pathway that involves a series of proteins in a protein kinase cascade in which each protein phosphorylates and activates the next protein in the pathway (A -> B...
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