Rapid generation of directed and unmarked deletions in Xanthomonas.
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.
Kamoun
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S
Department of Plant Pathology, University of California, Davis 95616.
Tola
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E
Kamdar
H
H
Kado
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CI
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*
D006602:Hexosyltransferases; C021513:levansucrase