Determination of synthesis rate and lifetime of bacterial mRNAs

Abstract

A method has been developed to determine the synthesis rate and lifetime of bacterial mRNAs, either bulk mRNA or specific mRNAs, with a minimum of physiological disturbance. The method uses hybridization of pulse-labeled RNA to specific probes followed by an evaluation based on a computer simulation of the labeling kinetics of different classes of RNA. The method was applied to the determination of bulk mRNA in Escherichia coli growing in glucose minimal medium: 60% of the instantaneous rate of RNA synthesis, or 2.3% of the total amount of RNA, was found to be mRNA with an average lifetime of 1.0 +/- 0.2 min (= 0.7 min half-life).