10 - MIC170 4/30/09 1 Lecture 10 (Thursday, 4/30) Reverse...

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Unformatted text preview: MIC170 4/30/09 1 Lecture 10 (Thursday, 4/30) Reverse genetics Gene phenotype (forward genetics: phenotype gene) When you want to study function of a certain gene/protein using yeast genetics; Starting from a gene rather than from a mutation of unknown gene 1. Identify a gene of interest in yeast Database search --- look for yeast genes that show high similarity to known gene 2. Gene disruption ---construct the null mutant of the gene Knockout one of the loci in diploid sporulation/germination examine the phenotype of haploid segregants phenotype provides a clue about the cellular function of the disrupted gene (see the figure below) Gene disruption: Gene A LEU2 a / leu2/leu2 diploid transfect Genomic DNA fragment containing gene A Replace the open reading frame with a selectable marker gene MIC170 4/30/09 2 Meiosis & sporulation leu- segregants (gene A is not disrupted) Leu + segregants (gene A null mutant) Homologous recombination MIC170 4/30/09...
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This note was uploaded on 06/11/2009 for the course MIC 170 taught by Professor Shiozaki during the Spring '09 term at UC Davis.

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10 - MIC170 4/30/09 1 Lecture 10 (Thursday, 4/30) Reverse...

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