ArticleBMC microbiology2010
A rapid and simple method for constructing stable mutants of Acinetobacter baumannii.
Article in BMC microbiology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 69 papers.
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Who cites it
69 citing papers in PubMed.
- Role of the AmvAR efflux system on the pathogenesis of Acinetobacter baumannii.Scientific reports · 2026Article
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- A novel GntR-ABC efflux system mediates oxidative stress response, drug resistance, motility and virulence inFrontiers in microbiology · 2026Article
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- The novel MFS efflux pump SxtP, regulated by the LysR-type transcriptional activator SxtR, is involved in the susceptibility to sulfamethoxazole/trimethoprim (SXT) and the pathogenesis ofAntimicrobial agents and chemotherapy · 2024Article
- Structure of a Rhs effector clade domain provides mechanistic insights into type VI secretion system toxin delivery.Nature communications · 2024Article
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- PMCNA_RS00975 activates NF-κB and ERK1/2 through TLR2 and contributes to the virulence ofFrontiers in cellular and infection microbiology · 2024Article
- AspFlex: Molecular Tools to Study Gene Expression and Regulation inACS synthetic biology · 2023Article
- A high-efficiency scar-free genome-editing toolkit for Acinetobacter baumannii.The Journal of antimicrobial chemotherapy · 2022Article
- The DdrR Coregulator of the Acinetobacter baumannii Mutagenic DNA Damage Response Potentiates UmuDAb Repression of Error-Prone Polymerases.Journal of bacteriology · 2022Article
- Polyphosphate Kinase Is Required for the Processes of Virulence and Persistence in Acinetobacter baumannii.Microbiology spectrum · 2022Article
- Identification of Genes inPlants (Basel, Switzerland) · 2022Article
- Effects of Inactivation of d,d-Transpeptidases of Acinetobacter baumannii on Bacterial Growth and Susceptibility to β-Lactam Antibiotics.Antimicrobial agents and chemotherapy · 2022Article
- Article
- The Acinetobacter baumannii disinfectant resistance protein, AmvA, is a spermidine and spermine efflux pump.Communications biology · 2021Article
- Role of Iron-Containing Alcohol Dehydrogenases inInternational journal of molecular sciences · 2021Article
- Role of AmpG in the resistance to β-lactam agents, including cephalosporins and carbapenems: candidate for a novel antimicrobial target.Annals of clinical microbiology and antimicrobials · 2021Article
- Phosphorylation of Extracellular Proteins inFrontiers in microbiology · 2021Article
- Direct interaction between RecA and a CheW-like protein is required for surface-associated motility, chemotaxis and the full virulence ofVirulence · 2020Article
9 more citing papers are in PubMed but not listed here.
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Authors and funding
8 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundAcinetobacter baumannii is a multidrug-resistant bacterium responsible for nosocomial infections in hospitals worldwide. Study of mutant phenotypes is fundamental for understanding gene function. The methodologies developed to inactivate A. baumannii genes are complicated and time-consuming; sometimes result in unstable mutants, and do not enable construction of double (or more) gene knockout mutant strains of A. baumannii.
resultsWe describe here a rapid and simple method of obtaining A. baumannii mutants by gene replacement via double crossover recombination, by use of a PCR product that carries an antibiotic resistance cassette flanked by regions homologous to the target locus. To demonstrate the reproducibility of the approach, we produced mutants of three different chromosomal genes (omp33, oxyR, and soxR) by this method. In addition, we disrupted one of these genes (omp33) by integration of a plasmid into the chromosome by single crossover recombination, the most widely used method of obtaining A. baumannii mutants. Comparison of the different techniques revealed absolute stability when the gene was replaced by a double recombination event, whereas up to 40% of the population reverted to wild-type when the plasmid was disrupting the target gene after 10 passages in broth without selective pressure. Moreover, we demonstrate that the combination of both gene disruption and gene replacement techniques is an easy and useful procedure for obtaining double gene knockout mutants in A. baumannii.
conclusionsThis study provides a rapid and simple method of obtaining stable mutants of A. baumannii free of foreign plasmidic DNA, which does not require cloning steps, and enables construction of multiple gene knockout mutants.
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