Hypo-osmotic shock - ppt1 and wildtype. Saccharomyces cerevisiae

Wild type and ppt1 mutant under hypo-osmotic shock. Cultures were grown with 1M sorbitol for ~20 hours, cells were collected by centrifugation and resuspended in YPD at time zero. Samples were collected at 0, 7, 15, 30 and 60 minutes after transfer to YPD. Experimental samples were used to generate Cy5-labeled cDNA probes, whereas mRNA reference pools extracted from cultures of the respective strains grown to early log phase under normal conditions, were used to generate Cy3-labeled cDNA probes. Cy5- and Cy3-labeled probes were hybridized together to microarrays printed with PCR-amplified fragments, representing 6280 of the Saccharomyces cerevisiae ORFs. Keywords: time-course

低渗胁迫下的野生型（Wild type）与ppt1突变体（ppt1 mutant）：将培养物在含1M山梨醇的培养基中培养约20小时，随后通过离心收集细胞，并于时间零点将其重悬于YPD培养基中。于转移至YPD培养基后的0、7、15、30与60分钟采集样品。实验样品用以制备Cy5标记的互补DNA (cDNA)探针；从在正常条件下培养至早期对数生长期的对应菌株培养物中提取的mRNA参考池，用以制备Cy3标记的cDNA探针。将Cy5与Cy3标记的探针共同杂交至以聚合酶链式反应（Polymerase Chain Reaction，简称PCR）扩增片段制备的微阵列上，该微阵列覆盖酿酒酵母（Saccharomyces cerevisiae）的6280个开放阅读框（Open Reading Frames，简称ORFs）。关键词：时间进程（time-course）