酿酒酵母中与钙离子稳态调控因子 ScRCH1细胞质膜定位相关的转录因子基因的筛选检测

摘要

Gene of ymr034c in Saccharomyces cerevisiae is homologous to the gene of Carch1 in Candida albicans gene,CaRCH1 and C. albicans were related to calcium ion,lithium ion,and nidazoles resistance. Therefore,it was named ymr034c as Scrch1. The earlier study showed that ScRCH1 localizes at plasma membrane under the coercion of high levels of extracellular calcium ion. In order to study on regulation mechanism of the expression of Scrch1 gene, single-gene deletion mutants for 223 of coding transcription factor in S. cerevisiae was screened through fluorescent mi-croscopy. The results showed that after calcium ion treatment,five deletion transcription genes of ngg1,hal9,crz1, ada2,and swi6,caused ScRCH1-GFP fail to localize at the plasma membrane. In addition,the deletion of ino2 gene led ScRCH1-GFP localized to the plasma membrane under the conditions even without the treatment of calcium ion.%酿酒酵母细胞中 ymr034c 基因与白念珠菌的 Carch1基因同源，CaRCH1与白念珠菌对钙离子、锂离子和硝唑类药物的耐受性相关。因此，把 ymr034c 命名为 Scrch1。前期研究结果显示，在胞外高钙离子胁迫条件下，ScRCH1定位于细胞质膜上。为了研究 Scrch1基因表达的调控机理，通过荧光显微镜技术对酵母细胞基因组中编码转录因子的223个单基因缺失菌株进行了筛选，分别检测了 ScRCH1-GFP 融合蛋白在它们中的亚细胞定位情况。结果发现，钙离子处理后，ngg1、hal9、crz1、ada2和 swi6五个转录因子基因的缺失造成ScRCH1-GFP 没有细胞质膜定位，而 ino2基因的缺失导致 ScRCH1-GFP 在不经钙离子处理的条件下即定位到细胞质膜上。