Chapter 4 Analyzing the Regulation of Antibiotic Production in Streptomycetes

Abstract

This chapter outlines the approaches and techniques that can be used to analyze the regulation of antibiotic production in streptomycetes. It describes how to isolate antibiotic nonproducing and overproducing mutants by UV, nitrosoguanidine (NTG), transposon, and insertion mutagenesis, and then how to use those mutants to identify regulatory genes. Other approaches to identify both pathway-specific and pleiotropic regulatory genes include overexpression and genome scanning. A variety of methods used to characterize pathway-specific regulatory genes for antibiotic biosynthesis are then covered, including transcriptional analysis and techniques that can be used to distinguish between direct and indirect regulation. Finally, genome-wide approaches that can be taken to characterize pleiotropic regulatory genes, including microarray and ChIP-on-Chip technologies, are described.