THY.AP4 Chemically Competent Cell

THY.AP4 Chemically Competent Cell

The Split Ubiquitin System (mbSUS) is a detection technology specifically designed to screen for interactions between transmembrane proteins. It uses a separated ubiquitin system to directly detect interactions between membrane proteins in their native state through a yeast two-hybrid system. This system employs the THY.AP4 yeast strain, which can directly transform plasmids for protein interaction validation or library screening experiments. The genotype of this strain is: MATa; ade2-, his3-, leu2-, trp1-, ura3-; lexA::ADE2, lexA::HIS3, lexA::lacZ. The first step involves selective growth screening using nutritional deficiency reporter genes (HIS3, ADE2), followed by β-galactosidase analysis using the LacZ reporter gene for colorimetric quantitative or semi-quantitative screening. There are three independent reporter genes regulated by different promoters to reduce the likelihood of false positives. The plasmids that can be used in this system include: pNXgate32-3HA, pXNgate21-3HA, and pMetYCgate. THY.AP4 competent cells are produced by special methods and can be stored at -80°C for 12 months, with a transformation efficiency of the pGADT7 plasmid detecting >10⁴ cfu/μg DNA.

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