EBY.VW4000 Chemically Competent Cell

EBY.VW4000Chemically Competent Cell

The EBY.VW4000 strain is derived from the CEN.PK2-1C yeast strain, which is of MATa type, with a transformation marker of ura3, making it suitable for selecting plasmids marked with URA3 for transformation, such as pYES2/NT, pYC2/NT, pDR196, p416GDP, pESC-URA, and others. The baker's yeast contains a large number of strictly regulated hexose transporters with various characteristics, which have the potential to transport glucose. Most yeasts contain 17 hexose transporters (HXT1 - HXT17), while CEN.PK2-1C only contains 16 hexose transporters (HXT1 - HXT16). GAL2, a galactose permease gene, is essential for the utilization of galactose and also has the ability to transport glucose. There are also three members of the maltose transporter family (Agt1, Ydl247, Yjr160) that can transport hexoses. Additionally, STL1 (Sugar Transporter-Like protein) is a proton symporter at the plasma membrane and is also thought to be involved in the glucose transport metabolism process. To eliminate the glucose uptake ability of CEN.PK2-1C yeast, the Cre/LoxP system was used to delete all 21 genes that can transport hexoses, including HXT1 - HXT16, GAL2, AGT1, YDL247w, and YJR160c, resulting in EBY.VW4000. As a hexose transporter-deficient yeast strain, EBY.VW4000's glucose absorption and transport activity is completely eliminated, and its capacity for absorbing and transporting other hexoses is also eliminated, making it an efficient experimental platform for discovering and studying natural or heterologous (from other species) hexose transporters. EBY.VW4000 cannot assimilate glucose or other hexoses as a carbon source, but it grows normally on media containing 2% maltose. For culturing WT cells, YPM medium is recommended; for selecting yeast cells containing plasmids, SM medium is recommended. The competent cells of EBY.VW4000 are prepared using a special process, can be stored at -80°C for 12 months, and the transformation efficiency of the pYES2 plasmid (5857bp, Amp^R) is >10⁴ cfu/μg DNA.

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