Latest Questions

(1) HIS3. Y2HGold cannot synthesize histidine and therefore cannot grow on media lacking this essential amino acid. When bait and prey proteins interact, the expression of the Gal4-responsive His3 gene allows cells to biosynthesize histidine and grow on minimal medium. 3-AT can be added to suppress background.
(2) ADE2. Y2HGold also cannot grow on minimal medium lacking adenine. However, when two proteins interact, Ade2 expression is activated, enabling these cells to grow on Ade- minimal medium.
(3) MEL1. MEL1 codes for α-galactosidase, an enzyme naturally present in many yeasts. Due to the two-hybrid interaction, α-galactosidase (MEL1) is expressed and secreted by the yeast cells. Yeast colonies expressing Mel1 turn blue in the presence of the chromogenic substrate X-α-Gal.
(4) AUR1-C. a dominant mutant version of the AUR1 gene, encodes inositol phosphorylceramide synthase. *AUR1-C* is expressed in the Y2HGold yeast strain due to protein-protein interaction bringing the GAL4 transcription activation and DNA-binding domains into proximity. AbA can be added for background suppression.The pABAi vector carries the Ura3 gene. The pGADT7 vector carries the Leu gene. The pGBKT7 vector carries the Trp gene.

Plates used for screening:

Single Dropout: SD/-Ura, SD/-Leu /+AbA

Double Dropout: DDO [SD/-Leu/-Trp], DDO/X [SD/-Leu/-Trp/X-α-Gal]

Triple Dropout: TDO [SD/-Leu/-Trp/-His]

Quadruple Dropout: QDO [SD/-Leu/-Trp/-His/-Ade]

The main service scope is as follows:

1、Nuclear Yeast One/Two-Hybrid library construction;

2、Membrane Yeast Two-Hybrid library construction;

3、Nuclear Yeast One/Two-Hybrid library screening;

4、Membrane Yeast Two-Hybrid library screening;

5、Nuclear Yeast One/Two-Hybrid point-to-point verification;

6、Membrane Yeast Two-Hybrid point-to-point verification