wsc1 mutant 3 hour exposure to zymolyase. Saccharomyces cerevisiae

We did transcription profiling on the effect of wsc1 deletion, gene involved in cell osmotic stress and cell wall stress response Keywords: cell wall stress response Overall design: S. cerevisiae (wsc1 mutant) was grown on YEPD. For Zymolyase experiments, yeast cells were grown overnight at 24 °C to an optical density 0.8 - 1 (A600). The culture was refreshed to 0.2 O.D and grown at 24 °C for 2h 30min. Next, culture was divided into two parts. One continues growing under same conditions (non-treated culture) while the other was supplemented with 0.8 units/ml Zymolyase 100T. Cells were collected at 3 hours of growth, frozen at -80 °C and processed for RNA extraction. The total RNA from two different cultures was analyzed, and, in addition, for each sample two different hybridizations were performed, including fluorochrome swapping in order to minimize transcriptional changes due to technical variability. This therefore corresponded to four DNA microarrays analyzed for each condition.

本研究针对参与细胞渗透压应激与细胞壁应激应答的WSC1基因的缺失效应开展了转录谱分析（transcription profiling）。
关键词：细胞壁应激应答
实验设计：以YEPD培养基培养wsc1缺失突变型酿酒酵母（S. cerevisiae）。
溶壁酶（Zymolyase）处理实验中，酵母细胞于24℃过夜培养至A600光密度值为0.8~1。将培养液稀释至初始光密度0.2，于24℃继续培养2小时30分钟。随后将培养液均分为两组：一组在相同条件下持续培养（未处理对照组），另一组添加终浓度0.8单位/毫升的溶壁酶100T（Zymolyase 100T）。培养3小时后收集细胞，置于-80℃冻存并进行RNA提取。
对两组独立培养物的总RNA进行分析；此外，每个样本均开展两次杂交实验，包含荧光染料互换实验，以最小化技术变异导致的转录组变化。因此每种培养条件对应四次DNA微阵列（DNA microarray）检测分析。