Chromosome Editing Using the Lambda Red Recombinase System in Chlamydia trachomatis and Chlamydia muridarum

This chapter describes a lambda Red-based recombineering approach for targeted genetic manipulation in Chlamydia species. The method uses a non-replicative plasmid encoding lambda Red recombination proteins together with locus-specific homology arms to generate precise gene deletions in the Chlamydia chromosome. Detailed protocols are provided for construct design, host cell-based selection, and genetic validation of recombinant strains, and noteworthy considerations for transforming different Chlamydia species and strains are given.