Yeast two-hybrid method

TL;DR: In this paper, a yeast two-hybrid method is described, which is characterized by comprising the steps that 1, library yeast is activated, and an AD-Y fusion vector is transformed intothe library yeast; 2, bait plasmids BD-X are transformed into yeast cells Y2H Gold; 3, recombined yeast of BD X and recombined Yeast containing the AD Y are fused into a diploid, the di-ploid yeast iscultured on an amplification culture medium containing zinc ions to obtain a bacterial colony, and then positive clone screening

Abstract: The invention provides a yeast two-hybrid method. The yeast two-hybrid method is characterized by comprising the steps that 1, library yeast is activated, and an AD-Y fusion vector is transformed intothe library yeast; 2, bait plasmids BD-X are transformed into yeast cells Y2H Gold; 3, recombined yeast of BD-X and recombined yeast containing the AD-Y are fused into a diploid, the diploid yeast iscultured on an amplification culture medium containing zinc ions to obtain a bacterial colony, and then positive clone screening is conducted. GAL4 protein in the yeast cells Y2H Gold used in the method is a dependent transcription factor of the zinc ions, therefore, the zinc ions are added into the amplification culture medium used in the method, interactions between low-abundance proteins are improved, the detection rate of low-interaction proteins is improved, and the positive clone rate is improved.