In Vivo Excision, Cloning, and Broad-Host-Range Transfer of Large Bacterial DNA Segments Using VEX-Capture

The performance of many bacterial genetic experiments would benefit from a convenient method to clone large sets of genes (20–100+kb) and transfer these genes to a wide range of other bacterial recipients. The VEX-Capture technique allows such large genomic segments to be cloned in vivo onto a broad-host-range IncP plasmid that is able to self-transfer to a wide variety of Gram-negative bacteria. The advantages of VEX-Capture are its efficiency, specificity, and use of common molecular biological techniques that do not require non-standard equipment and are easily applicable to many types of bacterial species. Here, we describe the VEX-Capture experimental protocol using Salmonella typhimurium as the source of the target DNA segment.