Random mutagenesis strategies for Campylobacter and Helicobacter species

Campylobacter and Helicobacter species are important pathogens in man and animals. The study of their virulence and physiology has been difficult due to the lack of tractable genetic tools, since many of the techniques established in Escherichia coli and related species were found to be non-functional in Campylobacter and Helicobacter species. The advent of functional genomics techniques in the last decade has been accompanied by the development of genetic tools, which take advantage of specific features of Campylobacter and Helicobacter, like natural transformation. This has allowed for the construction of random mutant libraries based on in vitro transposition or ligated loops followed by natural transformation and recombination, thus circumventing selection against sequences when cloning or passaging libraries through E. coli. Uses of the techniques have been in the study of motility, gene expression, and gene essentiality. In this chapter, we discuss the approaches and techniques used for the construction of random mutant libraries in both Campylobacter and Helicobacter.