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Team:uOttawa/Results
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'''Characterization of BBa_K642000 and BBa_K642004''' | '''Characterization of BBa_K642000 and BBa_K642004''' | ||
| - | We quantified the fluorescence of two tagged repressors: BBa_K642000 and BBa_K642004 by cloning them downstream of the yeast Gal10/1 promoter. The construct was then integrated using KanMX6 selection into the Ade2 locus of a BY4742 strain of S.cerevisiae. The resulting strain was innoculated in 3 mL of 1x SM + 2% Galactose + 2% Adenine overnight. The overnight culture was reinoculated into 1 mL of the same media at an OD600 of 0.02. After three hours the strain was analyzed for BFP fluorescence on a Cyan ADP flow cytometer. | + | We quantified the fluorescence of two tagged repressors: BBa_K642000 and BBa_K642004 by cloning them downstream of the yeast Gal10/1 promoter. The construct was then integrated using KanMX6 selection into the Ade2 locus of a BY4742 strain of ''S. cerevisiae''. The resulting strain was innoculated in 3 mL of 1x SM + 2% Galactose + 2% Adenine overnight. The overnight culture was reinoculated into 1 mL of the same media at an OD600 of 0.02. After three hours the strain was analyzed for BFP fluorescence on a Cyan ADP flow cytometer. |
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| + | [[File:BFP_graph.jpg]] | ||
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| + | Figure 1: In vivo BFP fluorescence of BBa_K642000, a TetR repressor tagged with yeast codon optimized BFP and BBa_K642004, a TetR repressor tagged with yeast codon optimized BFP and VP16 activation domain. These parts were integrated using KanMX6 selection into a BY4742 strain of ''S. cerevisiae''. | ||
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| + | [[File:Uninduced_WT.jpg]] | ||
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| + | Figure 2: Histogram of uninduced wildtype BY4742 | ||
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| + | [[File:Uninduced_tetr-bfp.jpg]] | ||
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| + | Figure 3: Histogram of uninduced BBa_K642000 -TetR-BFP | ||
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{{Template:uOttawa_Footer}} | {{Template:uOttawa_Footer}} | ||
Revision as of 02:40, 29 September 2011
Results
BioBrick characterization
Characterization of BBa_K642000 and BBa_K642004
We quantified the fluorescence of two tagged repressors: BBa_K642000 and BBa_K642004 by cloning them downstream of the yeast Gal10/1 promoter. The construct was then integrated using KanMX6 selection into the Ade2 locus of a BY4742 strain of S. cerevisiae. The resulting strain was innoculated in 3 mL of 1x SM + 2% Galactose + 2% Adenine overnight. The overnight culture was reinoculated into 1 mL of the same media at an OD600 of 0.02. After three hours the strain was analyzed for BFP fluorescence on a Cyan ADP flow cytometer.
Figure 1: In vivo BFP fluorescence of BBa_K642000, a TetR repressor tagged with yeast codon optimized BFP and BBa_K642004, a TetR repressor tagged with yeast codon optimized BFP and VP16 activation domain. These parts were integrated using KanMX6 selection into a BY4742 strain of S. cerevisiae.
Figure 2: Histogram of uninduced wildtype BY4742
Figure 3: Histogram of uninduced BBa_K642000 -TetR-BFP
