An improved counterselection cassette for use in Haemophilus influenzae.
Counterselectable cassettes are extremely useful in molecular biology and allow for the creation of unmarked deletion mutants or the introduction of point mutations. I have constructed an inducible sacB cassette, using the tetracycline repressor. When used in tandem with a kanamycin-resistance marker, the cassette was successful in creating unmarked mutants in Haemophilus influenzae. The inducible nature of the cassette avoids some of the common problems associated with the utilization of sacB in counterselection.
Department of Microbiology, Immunology, & Molecular Genetics, University of Kentucky, MS-423 Chandler Medical Center, Lexington, KY 40536-0298, USA. jason.johnston@uky.edu
Jason W
JW
Johnston
D001412:Bacillus subtilis / Q000235:genetics; D005798:Genes, Bacterial; D005819:Genetic Markers; D006193:Haemophilus influenzae / Q000235:genetics*; D006602:Hexosyltransferases / Q000235:genetics; D007613:Kanamycin Resistance / Q000235:genetics; D016254:Mutagenesis, Insertional; D009154:Mutation
D005819:Genetic Markers; D006602:Hexosyltransferases; C021513:levansucrase