RE&Complementation Lab Report

Met ura glu baseline transcription was observed b met

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+met -ura +glu, baseline transcription was observed B. -met -ura +gal, no significant growth was observed C. -met -ura +glu, no significant growth was observed. DEF. Yeast transformed with pYES2.1+ Met3 were plated on D. +met -ura +glu, significant growth was observed E. -met -ura +gal, significant growth was observed F. -met –ura +glu, no significant growth was observed. GHI. Yeast transformed with pBG1805+ MET3 were plated on G. +met -ura +glu, significant growth was
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observed H. -met -ura +gal, significant growth was observed I. -met –ura +glu, no significant growth was observed. Table 3: Complementation of met genes by yeast overexpression plasmids S. cerevisae transformed with pYES2.1- lacZ S. cerevisae transformed with pYES2.1+ Met3 S. cerevisae transformed with pBG1805+ MET3 +met -ura +glu Baseline growth Significant growth Significant growth -met -ura +gal No growth Significant growth Significant growth -met -ura +glu No growth No growth No growth Conclusions Observed band sizes for plasmid 4 match the expected band sizes for pYES2.1- lacZ , suggesting plasmid 4 is pYES2.1- lacZ . Observed band sizes for plasmid 10 match the expected band sizes for pYES2.1+ Met3 , suggesting plasmid 10 is pYES2.1+ Met3 . Observed band sizes for plasmid 14 match the expected band sizes for pBG1805+ MET3 , suggesting plasmid 14 is pBG1805+ MET3 . S.cerevisiae yeast transformed with pYES2.1- lacZ showed slight growth in repressed conditions of YC+met-ura+glu media, but not in YC+gal-ura-met, suggesting pYES2.1- lacZ does not complement the met3 auxotrophy. S.cerevisiae met3 auxotrophs grew in repressed conditions of YC+met-ura+glu media, and in YC+gal-ura-met after transformation with pYES2.1+ Met3 , suggesting that Met3 complements the met3 auxotrophy. S.cerevisiae met3 auxotrophs grew in repressed conditions of YC+met-ura+glu media, and in YC+gal-ura-met after transformation with pBG1805+ MET3 , suggesting that MET 3 complements the met3 auxotrophy. Citations
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O'Connor, Clare. Investigations in Molecular Cell Biology . Plymouth: Hayden-McNeil, 2012. Print. Amberg, D.C., Burke, D.J., and Strathern, J.N. (2005). Methods in Yeast Genetics (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY). P. 121
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