Signature-tagged mutagenesis: technical advances in a negative selection method for virulence gene identification

Signature-tagged mutagenesis (STM) is a powerful negative selection method, predominantly used to identify the genes of a pathogen that are required for the successful colonization of an animal host. Since its first description a decade ago, STM has been applied to screen a vast amount of transposon insertion mutants in 31 bacterial species. This has led to the identification of over 1,700 bacterial genes that are involved in virulence. Despite the preservation of the basic design, the STM method has been developed further owing to recent advances including different designs of the signature-tags and profound changes in the mode of detection. These advances promoted substantially the application range and versatility of the STM method.