Dynamics of transcription driven by the tetA promoter, one event at a time, in live Escherichia coli cells.

ABSTRACT: In Escherichia coli, tetracycline prevents translation. When subject to tetracycline, E. coli express TetA to pump it out by a mechanism that is sensitive, while fairly independent of cellular metabolism. We constructed a target gene, PtetA-mRFP1-96BS, with a 96 MS2-GFP binding site array in a single-copy BAC vector, whose expression is controlled by the tetA promoter. We measured the in vivo kinetics of production of individual RNA molecules of the target gene as a function of inducer concentration and temperature. From the distributions of intervals between transcription events, we find that RNA production by PtetA is a sub-Poissonian process. Next, we infer the number and duration of the prominent sequential steps in transcription initiation by maximum likelihood estimation. Under full

SUBMITTER: Muthukrishnan AB 

PROVIDER: S-EPMC3458540 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature