A method for generating marker-less gene deletions in multidrug-resistant Acinetobacter baumannii.

ABSTRACT:

Background

Acinetobacter baumannii is an important nosocomial pathogen that has become increasingly resistant to multiple antibiotics. Genetic manipulation of MDR A. baumannii is useful especially for defining the contribution of each active efflux mechanism in multidrug resistance. Existing methods rely on the use of an antibiotic selection marker and are not suited for multiple gene deletions.

Results

A tellurite-resistant (sacB⁺, xylE⁺) suicide vector, pMo130-TelR, was created for deleting the adeFGH and adeIJK operons in two clinical MDR A. baumannii, DB and R2 from Singapore. Using a two-step selection, plasmid insertion recombinants (first-crossover) were selected for tellurite resistance and the deletion mutants (second-crossover) were then selected for loss of sacB.

SUBMITTER: Amin IM 

PROVIDER: S-EPMC3717142 | biostudies-literature | 2013 Jul

REPOSITORIES: biostudies-literature