Differential expression analysis of T. thermophilus HB8 TTHA0995-deficient strain in response to iron. Thermus thermophilus HB8

We observed the expression profile of the total mRNA in Thermus thermophilus HB8 TTHA0995 deficient strain at 0 and 30 min after the addition of FeSO4. Overall design: Three TTHA0995 deficient strain of T. thermophilus HB8 were pre-cultured at 70 degC for 16 hours in 3 mL of TT medium containing 0.8% polypeptone, 0.4% yeast extract, 0.2% NaCl, 0.4 mM CaCl2, and 0.4 mM MgCl2, which was adjusted to pH 7.2 with NaOH. The cells (2 mL) were inoculated into 1 L of the same medium and then cultivated at 70°C for 5.5 hours (A600 value being ~1.53). Then the 1.0 mM of FeSO4 was added into the medium, and continued cultivation. Cells were collected at 0 and 30 min time point after the addition of iron ion, and then crude RNA was extracted. The expression of each mRNA at each time point was analyzed on a GeneChip as described under Sample Description Sheet of each sample.

我们检测了嗜热栖热菌（Thermus thermophilus）HB8的TTHA0995缺陷菌株在添加硫酸亚铁（FeSO4）后0分钟与30分钟时的总mRNA表达谱。
整体实验设计：将3株嗜热栖热菌HB8 TTHA0995缺陷菌株接种于3 mL含0.8%蛋白胨、0.4%酵母提取物、0.2%氯化钠、0.4 mM氯化钙及0.4 mM氯化镁的TT培养基（经NaOH调至pH 7.2）中，于70℃预培养16小时。取2 mL菌液接种至1 L相同培养基中，于70℃培养5.5小时（A600值约为1.53）。随后向培养基中加入终浓度1.0 mM的硫酸亚铁，继续培养。分别在添加铁离子后的0分钟和30分钟收集菌体，提取粗总RNA。参照各样本的样本描述单所述方法，利用基因芯片（GeneChip）分析各时间点下各mRNA的表达水平。