Alternative name Y2HGold Yeast Strains,Y2HGold,Yeast Packing 100ul Storage at -20DEG C Strains Resistance Kan Culture Medium YPDA Condition 28DEG C ,under the aerobic conditions
Y2HGold Yeast Strains Information
Culture medium: YPDA Strain category: yeast Culture conditions: 28鈩? aerobic, YPDA Plasmids Transformation: electric shock Preservation method: 30% glycerin, -20鈩?br /> Basic application: used for yeast two hybrid
Y2HGold strain is a strain of yeast GAL4 double heterozygous experiment developed by Clontech company, MATa type, which can directly transform plasmids or interact with MAT alpha yeast strain Y187 through mating operation for protein interaction verification or sieving test. Transformation marker is: TRP1, leu2, the report base: AbAr, HIS3, ADE2, MEL1. The Y2HGold -GAL4 yeast two heterozygous system requires two kinds of plasmids to be used together: PGBKT7 and PGADT7. Screening marker plasmid PGBKT7 TRP1 for expression of DNA-BD (from yeast transcription factor GAL4N 1~ terminal 174 amino acids) and target protein (Bait) fusion protein; screening marker plasmid PGADT7 LEU for expression of AD (GAL4 ~881 C terminal 768 amino acids) and target protein (Prey) fusion protein. GAL4 system principle: a complete yeast transcription factor GAL4 can be divided into two independent domains: the DNA binding domain (DNA-BD) located at the 1 to 174 amino acid segment at the N end and the transcriptional activation domain (AD) located at the 768 ~881 position of C terminal. DNA-BD can identify and combine the upstream activation sequence UAS of the GAL4-responsive gene. AD can activate the downstream genes of UAS to transcribe. BD and AD alone can not activate transcription, but when the two are close, they present a complete GAL4 activity, making the transcriptional expression of the downstream gene of the promoter containing UAS. Under normal conditions, BD is combined with AD to detection of proteins were fused with BD and AD, the formation of the bait fusion protein (bait -BD) and prey fusion protein (prey-AD), if bait interacts with prey, BD and AD will make close to each other, to form a complete GAL4, leading to activation of transcription report gene. Y2HGold four report: AbAr, HIS3, ADE2 gene, MEL1, respectively, by three different promoters (G1, G2, M1), only GAL4 recognition of the 17bp core area of the three promoter, the rest are different, and greatly reduce the probability of occurrence of false positive yeast two hybrid. In addition, the new report gene AbAr has the advantage of lower background compared with the previous nutritional defect reporter gene, and it can also reduce the probability of the occurrence of yeast two hybrid false positive.