Transposon Mutagenesis of Listeria monocytogenes

Abstract

Listeria monocytogenes is a Gram-positive, facultative intracellular foodborne pathogen that enters the human digestive tract after the consumption of contaminated food. Much research has been done to understand the virulence factors of Listeria monocytogenes. One useful tool to study these virulence factors has been transposon mutagenesis. Many mutants can be generated at a time by performing high-throughput mutagenesis using transposons and later screening these mutants to identify features related to particular functions in the bacteria. Many transposon delivery systems are not ideal for transposon studies in Listeria monocytogenes, as the transposon system is too large, has lower transposition efficiency, and a high rate of plasmid retention. Therefore, a new mariner-based transposition system has been developed for Listeria monocytogenes. This system is an ideal high-throughput transposon mutagenesis as the rate of transposition is high and random, along with very low plasmid retention capacity.