Difference between revisions of "Team:Waterloo/Parts"

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<a href="" ng-click="active='Gal1-CFP NSC'">Gal1-CFP NSC</a>
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<a href="" ng-click="active='Gal1-CFP-NSC'">Gal1-CFP NSC</a>
 
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                                     The ADH1 and Gal1 fusion.
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                                     The Gal1 and GFP fusion.
 
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                                 <img src="https://static.igem.org/mediawiki/2016/0/0c/T--Waterloo--Cup1_GFP.png" class="wcontent-img-solo37">
 
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                                     The ADH1 and Cup1 fusion.
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                                     The Cup1 and GFP fusion.
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Gal1-CFP NSC
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        <p class="paragraph-medium">This biobrick is a fusion of the Gal1,10 promoter, Cyan Fluorescent Protein (CFP), and the chaperone protein Hsp104.</p>
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                                <img src="https://static.igem.org/mediawiki/2016/0/05/T--Waterloo--Gal1_CFP_NSC.png" class="wcontent-img-solo37">
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                                <div class="wcontent-caption paragraph-medium">
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                                    The Gal1, CFP, and Hsp104 fusion.
 
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Revision as of 20:33, 15 October 2016

Our System
Hsp104-NSC
Hsp104-SC1
Hsp104-SC2
Biobrick
ADH1-GFP
Gal1-GFP
Cup1-GFP
Gal1-CFP NSC
Gal1-CFP SC1
Gal1-CFP SC2
Promoter Testing
Hsp104-Gal1
Hsp104-Cup1
pXP218-ADH1
pXP218-Gal1
pXP218-Cup1
Hsp104-NSC

This construct is a control to test the metabolic load of the Hsp104-CFP fusion protein and as a baseline against which we compare the change in [PSI+]/[psi-] state. We synthesized a fragment to clone into the [Hsp-PRS315]. This plasmid will be referred to as the Hsp104 plasmid and is used in several of our experiments.

The Hsp104 plasmid.
The insertion of a Gal1,10 promoter and CFP fusion into the Hsp104 plasmid.
Hsp104-SC1

CFP was synthesized with a stop codon in the place of Tyr-39 (TAC -> TAG) and amplified with flanking ApaI and BamHI sites. The premature stop codon (before the CFP fluorophore) was expected to truncate the protein during normal transcription. Fluorimetry readings were then used to quantify the amount of read-through for the CFP-tagged Hsp104.

The insertion of a Gal1,10 promoter and CFP fusion containing a premature stop codon into the Hsp104 plasmid.
The exact location of the premature stop codon used to make Hsp104-SC1.
Hsp104-SC2

CFP was synthesized with a stop codon in the place of Val-22 (TCA -> TAG) and amplified with flanking ApaI and BamHI sites. The premature stop codon (before the CFP fluorophore) was expected to truncate the protein during normal transcription. Fluorimetry readings were then used to quantify the amount of read-through for the CFP-tagged Hsp104.

The insertion of a Gal1,10 promoter and CFP fusion containing a premature stop codon into the Hsp104 plasmid.
The exact location of the premature stop codon used to make Hsp104-SC1.
ADH1-GFP

This biobrick contains a fusion of the ADH1 promoter and GFP. ADH1 is a eukaryotic promoter; the presence of ethanol induces it. The fusion between it and GFP allows the strength of the promoter to be quantized in varying concentrations of ethanol using the fluorescence of the GFP.

The ADH1 and GFP fusion.
ADH1-GFP

This biobrick is a fusion of the Gal1 promoter and GFP protein. Gal1 is a eukaryotic promoter; the presence of galactose induces it. The fusion allows the strength of the promoter to be quantized in the presence of varying concentrations of galactose using the fluorescence of the GFP.

The Gal1 and GFP fusion.
Cup1-GFP

This biobrick is a fusion of the Cup1 promoter and GFP protein. Cup1 is a eukaryotic promoter; the presence of Cu2+ induces it. The fusion allows the strength of the promoter to be quantized in the presence of varying concentrations of copper ions using the fluorescence of the GFP.

The Cup1 and GFP fusion.
Gal1-CFP NSC

This biobrick is a fusion of the Gal1,10 promoter, Cyan Fluorescent Protein (CFP), and the chaperone protein Hsp104.

The Gal1, CFP, and Hsp104 fusion.