Abstract
Publisher Summary This chapter discusses the role of Rho's in transcription attenuation in the tna Operon of Escherichia coli (E. Coli). The chapter describes methods allowing experimental analysis of the role of the tna operon leader peptide in influencing whether or not Rho-mediated transcription termination will occur. Various E. coli strains have been described as sources of S-30 preparations; S-30s from some strains offer the advantage that they will permit the use of linear DNA templates. Transcription attenuation is advantageous as a regulatory strategy as it allows cellular components and events that do not participate directly in transcription initiation decisions to influence gene expression. Thus, RNA sequences and structures play a decisive role in the process of transcription attenuation. The most widely studied termination factor is the Rho protein of E. Coli. Rho-dependent transcription termination in the leader region of the tna operon can be readily demonstrated by comparing the products of two reactions, one with bicyclomycin addition (50 μg/ml), and the second without this antibiotic.
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