EGY48 Chemically Competent Cell

For the first use of Carrier DNA，boil the tube containing Carrier DNA in boiling water for 5 min，then place it on ice immediately. Store it at -20℃ for later use after application. Thaw Carrier DNA on ice before next use. Re-denature Carrier DNA after 3-5 repeated freeze-thaw cycles. Competent cells are best thawed on ice. Using high-concentration plasmids for transformation can correspondingly reduce the bacterial volume for final plating. Saccharomyces cerevisiae is temperature-sensitive with an optimal growth temperature of 28℃-30℃; temperatures above 31℃ will affect yeast growth. Yeast grows slower in dropout medium than in YPDA medium. The more nutritional defects the medium contains，the slower the yeast grows. EGY48 is a MATα haploid strain of Saccharomyces cerevisiae，specifically designed for the LexA heterologous yeast two-hybrid system. It is mainly used for the screening and verification of interactions among soluble nuclear proteins，with LEU2 as the primary nutritional reporter gene，which can be combined with the lacZ colorimetric reporter for dual confirmation. The transformation markers are his3，trp1 and ura3; the reporter gene is LEU2，and its upstream activation sequence is derived from LexA operator (x6)，which can only initiate the expression of LEU2 when bait and prey proteins interact with each other. The transformation efficiency of EGY48 competent cells detected with pGBKT7 plasmid (7303bp，Kan?) is higher than 10? cfu/μg DNA. Genotype: MATα，ura3，his3，trp1，LexAop (x6)-LEU2