EGY48 Chemically Competent Cell

EGY48 Chemically Competent Cell

The EGY48 strain is a yeast two-hybrid strain developed by Clontech using the LexA system. It is of the MATα type and can be directly transformed with plasmids for protein interaction validation or screening experiments. The transformation markers are: his3, trp1, ura3, and the reporter gene is: LEU2; the reporter gene UAS (Upstream Activation Sequence) comes from LexA op(x6), and LEU2 expression can only be initiated when Bait and Prey interact. The EGY48-LexA yeast two-hybrid system requires the use of three plasmids: pLexA, pB42AD, and p8op-LacZ. The selection marker for plasmid pLexA is HIS3, which is used to express a fusion protein of DNA-BD (composed of 202 amino acid residues from the prokaryotic LexA protein) and the target protein (Bait); the selection marker for plasmid pB42AD is TRP1, which is used to express a fusion protein of AD (composed of 88 amino acid residues from the herpes simplex virus B42AD protein) and the target protein (Prey); the selection marker for the reporter plasmid p8op-LacZ is URA3, and the reporter gene is LacZ. The reporter gene UAS comes from LexA op(x6), and LacZ expression can only be initiated when Bait and Prey interact. The EGY48 competent cells are produced through a special process and can be stored at -80°C for 12 months; the transformation efficiency of the pGBKT7 plasmid (7303 bp, Kan^R) is >10⁴ cfu/μg DNA.

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