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.
Wilson
James W
JW
Center for Infectious Diseases and Vaccinology, Biodesign Institute, Arizona State University, Tempe, USA.
Nickerson
Cheryl A
CA
D001431:Bacteriological Techniques; D003001:Cloning, Molecular / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D022761:Gene Transfer, Horizontal; D005821:Genetic Techniques; D005822:Genetic Vectors; D016680:Genome, Bacterial; D010957:Plasmids / Q000235:genetics; D012486:Salmonella typhimurium / Q000235:genetics
D004269:DNA, Bacterial