Y8930 strain酵母菌株 BioVector NTCC质粒载体菌种细胞基因保藏中心

Y8930 strain酵母菌株 BioVector NTCC质粒载体菌种细胞基因保藏中心 Y8930
Cat#：NTCC596123
Y8930 strain. 1Vial. Storage:4℃ S. cerevisiae strains Y8800 (MATa) and Y8930 (MATα) contain deletions of the GAL4 and GAL80 genes encoding Gal4p and its repressor Gal80p, respectively. In the absence of Gal80p, galactose is not required for activation of Gal4p-inducible promoters. The strains contain three Gal4p inducible reporter genes, providing four phenotypes to identify true interactors. The reporter genes are HIS3, providing for growth on plates lacking histidine, LacZ, for colorimetric detection of Gal4p activity, and ADE2, providing growth on media lacking adenine, as well as a range of color from red to white depending on the strength of Gal4p expression. In addition, the strains are resistant to cycloheximide, aiding in plasmid shuffling. Y8800 and Y8930 were generated by adding cycloheximide resistance to the PJ69-4 Y2H strains. PJ69-4 is described in P. James et. al. 1996. Y8800 / Y8930 Auxotrophic markers Description leu2-3,112 Need for Leu in medium trp1-901 Need for Trp in medium his3Δ200 Need for His in medium ade2-101 Need for Ade in medium ura3-52 Need for Ura in medium Other Description gal4Δ Deletion of endogenous Gal4p transcription factor gal80Δ Deletion of Gal4p repressor. cyh2R Resistance to Cycloheximide Reporters Description GAL2::ADE2 ade2-101 locus replaced by GAL2 promoter driven ADE2 GAL1::HIS3@LYS2 GAL1 promoter driven HIS3 gene downstream of LYS2 gene GAL7::LacZ@met2 GAL7 promoter driven LacZ gene inserted in the MET2 locus Recovery 1. Obtain an YPD plate. 2. Using a sterile pipette tip, touch the yeast growing within the punctured area of the stab culture. (A sterilized wire loop or sterile toothpick can be used in place of a sterile pipette tip.) 3. Run this tip lightly over a section of the plate, as shown in the figure, to create streak #1. 4. Using another sterile pipette tip, pass through streak #1 and spread the bacteria over a second section of the plate, to create streak #2. 5. Using a third sterile pipette tip, pass through streak #2 and spread the bacteria over the last section of the plate, to create streak #3. 6. Grow in a 30 o C incubator (unless a different growth temperature is indicated on the plasmid datasheet). 7. After 24-48h incubation, single colonies should be visible. If the yeast growth is too dense, re-streak onto a new YPD plate to obtain single colonies.
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