Difference between revisions of "Part:BBa K642007"
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We were able to characterize the expression of the Adh1 distal-Gal1 proximal promoter by cloning yeast enhanced GFP downstream of it. The promoter was integrated using NatMX selection into the Ade4 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 GFP fluorescence on a Cyan ADP flow cytometer. | We were able to characterize the expression of the Adh1 distal-Gal1 proximal promoter by cloning yeast enhanced GFP downstream of it. The promoter was integrated using NatMX selection into the Ade4 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 GFP fluorescence on a Cyan ADP flow cytometer. | ||
| − | + | [[Image:Induced_WT-adh.jpg]] | |
Figure 1: Histogram of galactose induced wildtype BY4742 | Figure 1: Histogram of galactose induced wildtype BY4742 | ||
| − | + | [[Image:Induced_adh-distal-gal-proximal.jpg]] | |
Figure 2: Histogram of the galactose induced ADH1 distal-Gal1 proximal promoter with GFP cloned downstream | Figure 2: Histogram of the galactose induced ADH1 distal-Gal1 proximal promoter with GFP cloned downstream | ||
Revision as of 21:52, 24 October 2011
Constitutive Adh1 distal-Gal1 proximal promoter tagged with yeGFP
This is the distal region of the mid length Adh1 promoter (BBa_K319005) submitted last year by the uOttawa team fused to the proximal region of the Gal1 promoter. We have demonstrated that it is a functional chimeric promoter and is inducible in the presence of galactose. The promoter is also tagged with the reporter yeGFP (BBa_K319039) which was submitted by the uOttawa team last year.
Characterization of BBa_K642007
We were able to characterize the expression of the Adh1 distal-Gal1 proximal promoter by cloning yeast enhanced GFP downstream of it. The promoter was integrated using NatMX selection into the Ade4 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 GFP fluorescence on a Cyan ADP flow cytometer.
Figure 1: Histogram of galactose induced wildtype BY4742
File:Induced adh-distal-gal-proximal.jpg
Figure 2: Histogram of the galactose induced ADH1 distal-Gal1 proximal promoter with GFP cloned downstream
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 89
Illegal BsaI.rc site found at 1379