Density gradient enrichment of Escherichia coli conditional msbA mutants.
Insight into the mechanism of lipid transport to the outer membrane of gram-negative bacteria has been hampered by the lack of an effective genetic screen for defective mutants. This work demonstrates an enrichment of conditional mutants defective in lipopolysaccharide export by Ludox density gradient centrifugation and selection for detergent resistance. New temperature-sensitive mutants with lipid export defects were isolated with single missense mutations in msbA. The results demonstrate the power of this approach for the study of lipid export in Escherichia coli.
Doerrler
William T
WT
Department of Biological Sciences, Louisiana State University, 202 Life Sciences Building, Baton Rouge, LA 70803, USA. wdoerr@lsu.edu
D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D002499:Centrifugation, Density Gradient / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000378:metabolism; D050505:Mutant Proteins / Q000235:genetics; D020125:Mutation, Missense; D012822:Silicon Dioxide
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D008070:Lipopolysaccharides; C079726:MsbA protein, Bacteria; D050505:Mutant Proteins; C024922:Ludox AM; D012822:Silicon Dioxide