Chromosomal Complementation Using Tn7 Transposon Vectors in Enterobacteriaceae.
Abstract
Genetic complementation in many bacteria is commonly achieved by re-introducing functional copies of the mutated or deleted genes on a recombinant plasmid. Chromosomal integration systems using the Tn7 transposon have the advantage of providing a stable single-copy integration that does not require selective pressure. Previous Tn7 systems have been developed although none have been shown to work effectively in a variety of Enterobacteria. Herein, we have developed several mini-Tn7 and transposase vectors to provide a more versatile system. Transposition of Tn7 at the chromosomal attTn7 site was achieved by a classical conjugation approach, where the donor strain harbored the mini-Tn7 vector and the recipient strain possessed the transposase vector. This approach was efficient in 5 different pathogenic enterobacterial species. Thereby, 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 introduction of foreign genes of use for biotechnology applications, vaccine development, or for use in gene expression and gene fusion constructs.