Experiment 5 - 1 Chris Stanton April 16, 2009 Leah,...

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1 Chris Stanton April 16, 2009 Leah, Thursday Rm 41 Experiment 5: Complementation Analysis of Histidine Requiring Mutants of Saccharoyces cerevisiae Purpose: The purpose of the experiment was to mate MATa and MAT-alpha strains of yeast to observe complementation between mutations in four different genes coding for histidine synthesis in yeast. Complementation analysis is a functional test that helps to determine if two mutations are part of the same gene or not. If two mutants complement each other, otherwise known as forming a phenotypically wild-type diploid, the two mutations are in different genes or different protein coding sequences within the same gene, as in the HIS4 gene that is analyzed in this lab Controls: The controls in this experiment were the fact that all of the MATa haploid strains were leu - and all of the MAT-alpha strains were auxotrophs for his and another amino acid. By plating on the YPD plate, min + his plate, and min plate you are able to control for the auxotrophies of other amino acids. The leu 2 MATa cells needed to be mated to become heterozygous for the leu 2 gene in order to grow on min or min + his because if they were haploid, they would not be able to grow without the presence of leucine. Similarly, all of the MAT-alpha strains were auxotrophs for a different amino acid, so they could not grow as haploid strains on min or min + his. This assured that all the yeast that grew on the min plates resulted from MATa his - leu 2 mutants and MAT-alpha auxotroph his - mutants mating. On the YPD plate, growth was expected everywhere because it has all the substances yeast needs to live. The YPD plate served as a transfer
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Experiment 5 - 1 Chris Stanton April 16, 2009 Leah,...

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