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Updated: May 28, 2026

Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)
Published on: March 16, 2012
An improved counterselection cassette for use in Haemophilus influenzae
1Department of Microbiology, Immunology, & Molecular Genetics, University of Kentucky, MS-423 Chandler Medical Center, Lexington, KY 40536-0298, USA. jason.johnston@uky.edu
Abstract:
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.

