Markerless deletions of pil genes in Myxococcus xanthus generated by counterselection with the Bacillus subtilis sacB gene.
In-frame deletions of pilA and pilS were constructed in Myxococcus xanthus with a plasmid integration-excision strategy facilitated by sacB. sacB conferred sucrose sensitivity upon its M. xanthus host only when it lay in the same orientation as adjacent M. xanthus genes. Gene orientation also affected the efficiency of sucrose counterselection in the sucrose-sensitive strains. The deltapilA mutant lacked pili and social motility, while the deltapilS mutant showed no defect in either phenotype.
Wu
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Department of Biochemistry, Stanford University School of Medicine, California 94305, USA.
Kaiser
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D001412:Bacillus subtilis / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics*; D036781:Fimbriae Proteins; D017353:Gene Deletion; D005798:Genes, Bacterial; D006602:Hexosyltransferases; D016254:Mutagenesis, Insertional; D016941:Myxococcus xanthus / Q000235:genetics*; D012641:Selection, Genetic; D013395:Sucrose / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics*
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D014157:Transcription Factors; C080687:pilS protein, Bacteria; D036781:Fimbriae Proteins; D013395:Sucrose; D006602:Hexosyltransferases; C021513:levansucrase