Team:Waterloo/Parts
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.
