MudSacI, a transposon with strong selectable and counterselectable markers: use for rapid mapping of chromosomal mutations in Salmonella typhimurium.
The transposable bacteriophage Mu and its mini-Mu derivatives are useful tools for the genetic analysis of many bacteria. A variety of antibiotic-resistant Mu derivatives have been constructed, allowing direct selection for cells which contain the transposon. However, in many cases a counterselection against the transposon would greatly facilitate further genetic analysis. In this paper we report the construction of MudSacI, a mini-Mu derived transposon containing the sacB (secretory levansucrase) gene of Bacillus subtilis, which confers sucrose sensitivity upon gram-negative bacteria. We describe the use of this transposon as a tool for rapid genetic mapping of chromosomal genes in Salmonella typhimurium. Simple modifications of this approach should facilitate rapid mapping in many other bacteria as well.
Lawes
M
M
Department of Microbiology, University of Illinois, Urbana 61801.
Maloy
S
S
D010583:Bacteriophage mu / Q000235:genetics; D002874:Chromosome Mapping / Q000379:methods*; D002876:Chromosomes, Bacterial / Q000235:genetics*; D004251:DNA Transposable Elements / Q000235:genetics; D005816:Genetic Complementation Test; D005819:Genetic Markers; D006090:Gram-Negative Bacteria / Q000235:genetics; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D016254:Mutagenesis, Insertional; D009154:Mutation / Q000235:genetics*; D015183:Restriction Mapping; D012486:Salmonella typhimurium / Q000235:genetics*; D012641:Selection, Genetic; D013395:Sucrose / Q000633:toxicity
D004251:DNA Transposable Elements; D005819:Genetic Markers; D013395:Sucrose; D006602:Hexosyltransferases; C021513:levansucrase