Construction of Hexose Transporter-like HXT1 Deletion Mutant in Pichia pastoris

Abstract:

Glucose was transported by the large number of hexose transporters in yeast cells. There were 18 hexose transporter genes had been identified in Saccharomyces cerevisiae. However, as an excellent expression system, there was no information of these genes had been reported in Pichia pastoris. Based on high homologous recombination efficiency in yeast, we chose G418 resistance for screening, 200 bp were cloned from the up and down sequences of HXT1 ORF respectively, then ligated to the 5￠ and 3￠ end of G418 resistance gene for recombination. After electroporation of GS115 spheroplast and screened through different G418 concentration plates, finally we obtained one HXT1 gene deletion mutant named GS115ΔHXT1. The growth rate and glucose consumption of this mutant were both lower than the wide type.