mRNA degradation in Mycobacterium tuberculosis with DosR ectopically induced

After induction of DosR from a tet regulated promoter (uninduced controls were treated with an equivalent ammount of the solvent, DMSO), transcription was arrested using 50 ug/ul of rifampin leaving RNA degradation as the sole vector responsible for mRNA abundance. mRNA half-lives were calculated from the decay of signal intensity from T0. Three replicates for each condition (DosR induced and uninduced) were sampled after 0, 10, 20, 30, and 60 minutes of mRNA decay for a total of 30 arrays.

通过四环素（tet）调控的启动子诱导DosR表达后（未诱导对照组使用等体积溶剂二甲基亚砜（DMSO）处理），我们采用50 μg/μl的利福平阻断转录过程，使得RNA降解成为决定mRNA丰度的唯一因素。mRNA的半衰期通过以T0时刻为基准的信号强度衰减情况计算得到。针对DosR诱导组与未诱导组两种实验条件各设置三次生物学重复，分别在mRNA降解启动后的0、10、20、30和60分钟进行取样，最终共计完成30组基因微阵列实验。