title: Rapid generation of directed and unmarked deletions in Xanthomonas.
abstract: We have devised a rapid four-step procedure for the generation of directed and unmarked chromosomal deletions in bacteria, based on the use of a novel cloning vector containing the Bacillus subtilis sacB gene that encodes levansucrase and confers sucrose sensitivity, which can be used for counter-selection. Using this technique, we describe the construction of a 6.5 kb directed and unmarked deletion in a phytopathogenicity region of the chromosome in Xanthomonas campestris. This procedure allows rapid and easy transfer of a wide variety of mutant allelic DNA to the bacterial chromosome, and should be adaptable to various bacteria besides Xanthomonas spp.
authors.affiliation: Department of Plant Pathology, University of California, Davis 95616.
authors.forename: S
authors.initials: S
authors.lastname: Kamoun
authors.forename: E
authors.initials: E
authors.lastname: Tola
authors.forename: H
authors.initials: H
authors.lastname: Kamdar
authors.forename: C I
authors.initials: CI
authors.lastname: Kado
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D005821:Genetic Techniques; D006602:Hexosyltransferases / Q000235:genetics; D016296:Mutagenesis / Q000235:genetics; D010957:Plasmids / Q000502:physiology; D016959:Xanthomonas campestris / Q000235:genetics*
chemical_list: D006602:Hexosyltransferases; C021513:levansucrase