Construction of Hexose Transporter-like HXT1 Deletion Mutant in Pichia pastoris
- VernacularTitle:毕赤酵母己糖运载体HXT1基因缺失菌株的构建
- Author:
Wen-Wen ZHANG
;
Ping ZHANG
;
Yao-Ji XUAN
;
Xiang-Shan ZHOU
;
Yuan-Xing ZHANG
;
- Publication Type:Journal Article
- Keywords:
Pichia Pastoris,Hexose transporter,G418 resistance,Gene deletion
- From:
Microbiology
2008;0(09):-
- CountryChina
- Language:Chinese
-
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 resis-tance gene for recombination. After electroporation of GS115 spheroplast and screened through different G418 concentration plates,finally we obtained one HXT1 gene deletion mutant named GS115?HXT1. The growth rate and glucose consumption of this mutant were both lower than the wide type.
