Difference between revisions of "Team:Waterloo/Parts"
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</div> | </div> | ||
<img src="https://static.igem.org/mediawiki/2016/6/6c/T--Waterloo--CFP_SC1.png" class="wcontent-img-solo37"> | <img src="https://static.igem.org/mediawiki/2016/6/6c/T--Waterloo--CFP_SC1.png" class="wcontent-img-solo37"> | ||
| + | <div class="wcontent-caption paragraph-medium"> | ||
| + | The exact location of the premature stop codon used to make Hsp104-SC1. | ||
| + | </div> | ||
| + | </div> | ||
| + | </div> | ||
| + | </div> | ||
| + | <div class="col-lg-8"> | ||
| + | <div class="wcontent-box" id="SC2" ng-show="active == 'SC2'"> | ||
| + | <div class="wcontent-title"> | ||
| + | Hsp104-SC2 | ||
| + | </div> | ||
| + | <div class="wcontent"> | ||
| + | <p class="paragraph-medium">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.</p> | ||
| + | <img src="https://static.igem.org/mediawiki/2016/7/74/T--Waterloo--HSP104_LEU_Addgene_Annotate_and_CFP_Gal_Stop1_ApaBam.png" class="wcontent-img-solo37"> | ||
| + | <div class="wcontent-caption paragraph-medium"> | ||
| + | The insertion of a Gal1,10 promoter and CFP fusion containing a premature stop codon into the Hsp104 plasmid. | ||
| + | </div> | ||
| + | <img src="https://static.igem.org/mediawiki/2016/f/ff/T--Waterloo--CFP_SC2.png" class="wcontent-img-solo37"> | ||
<div class="wcontent-caption paragraph-medium"> | <div class="wcontent-caption paragraph-medium"> | ||
The exact location of the premature stop codon used to make Hsp104-SC1. | The exact location of the premature stop codon used to make Hsp104-SC1. | ||
Revision as of 20:22, 15 October 2016
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.
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.
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.
