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酿酒酵母转座标签插入突变体263-H9中高盐胁迫基因的确定

Identification of the Gene Correlated with Salt Stress in the Saccharomyces cerevisiae 263-H9 Mutant

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【作者】 于典科张小华刘向勇鲍晓明高东

【Author】 YU Dian-Ke, ZHANG Xiao-Hua, LIU Xiang-Yong, BAO Xiao-Ming, GAO Dong (State Key Laboratory of Microbial Technology, Shandong University, Ji’nan 250100, China)

【机构】 山东大学微生物技术国家重点实验室山东大学微生物技术国家重点实验室 济南250100济南250100

【摘要】 突变体263-H9是利用mTn3转座标签对酿酒酵母(Saccharomycescerevisiae)W303-1A诱变、筛选得到的。该突变体表现出对多种逆境胁迫(1.5mol/L山梨醇高渗透压胁迫、0.65mol/LNaCl高盐胁迫和15℃低温胁迫)敏感的表型特征,而且与其他突变体不同其转座标签的插入位点是GIP2和YER053C-A的基因间隔区域。本文通过基因敲除、基因组文库功能互补等多种分子生物学和遗传学方法,确定了突变体263-H9的敏感表型不是由于转座标签的插入直接引起的,而是盐胁迫反应信号传导途经中重要的基因PBS2发生部分缺失,造成该基因不能正常表达,而导致的表型变化。

【Abstract】 The mutant 263-H9 with hypersensitivity to several stress conditions (1.5 mol/L Sorbitol, 0.65 mol/L NaCl and 15℃) was obtained by using transposon mutagenesis in the Saccharomyces cerevisiae strain W303-1A. Unlike other mutants the transposon in 263-H9 was intergenic between GIP2 and YER053C-A. Using gene knockout, a yeast genomic library and other methods, the gene correlated with the salt stress response was identified. The data indicated that the phenotype of 263-H9 was not directly caused by the insertion of the transposon. On the other hand, the hypersensitivity to salt and other stress conditions was due to the deletion of 5 base pairs close to position 936bp in the PBS2 gene essential for HOG signal pathway regulation under salt stress.

【关键词】 转座标签酿酒酵母高盐胁迫PBS2
【Key words】 transposon taggingSaccharomyces cerevisiaesalt stressPBS2
【基金】 国家自然科学基金项目(编号:30170021,30570031);留学回国人员科研资助项目~~
  • 【分类号】Q933
  • 【被引频次】8
  • 【下载频次】200
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