Chromosomal Complementation Using Tn7Transposon Vectors in Enterobacteriaceae

Abstract

ABSTRACTGenetic complementation in many bacteria is commonly achieved by reintroducing functional copies of the mutated or deleted genes on a recombinant plasmid. Chromosomal integration systems using the Tn7transposon have the advantage of providing a stable single-copy integration that does not require selective pressure. Previous Tn7systems have been developed, although none have been shown to work effectively in a variety of enterobacteria. We have developed several mini-Tn7and transposase vectors to provide a more versatile system. Transposition of Tn7at the chromosomalattTn7site was achieved by a classical conjugation approach, wherein the donor strain harbored the mini-Tn7vector and the recipient strain possessed the transposase vector. This approach was efficient for five different pathogenic enterobacterial species. Thus, this system provides a useful tool for single-copy complementation at an episomal site for research in bacterial genetics and microbial pathogenesis. Furthermore, these vectors could also be used for the introduction of foreign genes for use in biotechnology applications, vaccine development, or gene expression and gene fusion constructs.