酿酒酵母TRK1和TRK2钾吸收缺陷型菌株的构建

The Defective Potassium Uptake by TRK1 and TRK2 Mutant of Saccharomyces cerevisiae Using Homologous Recombination

摘要:

为更好的进行钾素营养有关基因表达调控和功能性研究, 我们采用同源重组法通过重叠引物扩增分别将URA3和HIS3基因替代酿酒酵母的TRK1和TRK2基因, 并以酿酒酵母的尿嘧啶合成酶URA3基因和组氨酸合成酶HIS3为标记基因, 在不含尿嘧啶和组氨酸的基本培养基筛选转化子获得了钾离子转运蛋白TRK1和TRK2基因缺失的酿酒酵母钾素营养缺陷型菌株, 该菌株在低K+培养基中导入拟南芥K+转运体基因AtKuP1可恢复正常生长。

Abstract:

In order to perform the regulation and functional study and the screening of potassium uptake genes, the high affinity K+ uptake genes TRK1 and TRK2 in Saccharomyces cerevisiae were replaced with URA3 and HIS3 alternately using the methods of homologous recombination. URA3 and HIS3 of S. Cerevisiae were used as marker gene, the defective K+ uptake TRK1 and TRK2 mutant of S. Cerevisiae were selected in the basic medium without Histidine and Uracil.