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Characterization of the chromosomal aac(6')-Ij gene of Acinetobacter sp. 13 and the aac(6')-Ih plasmid gene of Acinetobacter baumannii.

ABSTRACT: The amikacin resistance genes aac(6')-Ih of Acinetobacter baumannii BM2686 and aac(6')-Ij of Acinetobacter sp. 13 BM2689 encoding aminoglycoside 6'-N-acetyltransferases were characterized. The 441-bp coding sequences predict proteins with calculated masses of 16,698 and 16,677 Da, respectively. Analysis of the deduced amino acid sequences indicated that the proteins belonged to a subfamily of 6'-aminoglycoside acetyltransferase type I enzymes from gram-negative bacteria. The aac(6')-Ih gene of BM2686 was located on a 13.7-kb nonconjugative plasmid. The aac(6')-Ij gene from BM2689 was not transferable either by conjugation to Escherichia coli or A. baumannii or by transformation to Acinetobacter calcoaceticus. Plasmid DNA from BM2689 did not hybridize with an intragenic aac(6')-Ij probe. These results suggest a chromosomal location for this gene. The aac(6')-Ij gene was detected by DNA hybridization in all 28 strains of Acinetobacter sp. 13 tested but not in other Acinetobacter strains, including A. baumannii, proteolytic genospecies 4, 6, 14, 15, 16, and 17, and ungrouped strains. The aac(6')-Ih and -Ij probes did not hybridize in dot blot assays with DNA from members of the families Enterobacteriaceae and Pseudomonadaceae that produced 6'-N-acetyltransferases. These data suggest that the genes are confined to the Acinetobacter genus and that the aac(6')-Ij gene is species specific and may be used to identify Acinetobacter sp. 13.

SUBMITTER: Lambert T

PROVIDER: S-EPMC284657 | biostudies-other | 1994 Sep

REPOSITORIES: biostudies-other

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Characterization of the chromosomal aac(6')-Ij gene of Acinetobacter sp. 13 and the aac(6')-Ih plasmid gene of Acinetobacter baumannii.

Lambert T T Gerbaud G G Courvalin P P

Antimicrobial agents and chemotherapy 19940901 9

The amikacin resistance genes aac(6')-Ih of Acinetobacter baumannii BM2686 and aac(6')-Ij of Acinetobacter sp. 13 BM2689 encoding aminoglycoside 6'-N-acetyltransferases were characterized. The 441-bp coding sequences predict proteins with calculated masses of 16,698 and 16,677 Da, respectively. Analysis of the deduced amino acid sequences indicated that the proteins belonged to a subfamily of 6'-aminoglycoside acetyltransferase type I enzymes from gram-negative bacteria. The aac(6')-Ih gene of B ...[more]

PMID: 7810994

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Project description:Rates of infection with hospital-acquired Acinetobacter baumannii have exploded over the past decade due to our inability to limit persistence and effectively treat disease. A. baumannii quickly acquires antibiotic resistance, and its genome encodes mechanisms to tolerate biocides and desiccation, which enhance its persistence in hospital settings. With depleted antibiotic options, new methods to treat A. baumannii infections are desperately needed. A comprehensive understanding detailing A. baumannii cellular factors that contribute to its resiliency at genetic and mechanistic levels is vital to the development of new treatment options. Tools to rapidly dissect the A. baumannii genome will facilitate this goal by quickly advancing our understanding of A. baumannii gene-phenotype relationships. We describe here a recombination-mediated genetic engineering (recombineering) system for targeted genome editing of A. baumannii. We have demonstrated that this system can perform directed mutagenesis on wide-ranging genes and operons and is functional in various strains of A. baumannii, indicating its broad application. We utilized this system to investigate key gene-phenotype relationships in A. baumannii biology important to infection and persistence in hospitals, including oxidative stress protection, biocide resistance mechanisms, and biofilm formation. In addition, we have demonstrated that both the formation and movement of type IV pili play an important role in A. baumannii biofilm. Importance: Acinetobacter baumannii is the causative agent of hospital-acquired infections, including pneumonia and serious blood and wound infections. A. baumannii is an emerging pathogen and has become a threat to public health because it quickly develops antibiotic resistance, making treatment difficult or impossible. While the threat of A. baumannii is well recognized, our understanding of even its most basic biology lags behind. Analysis of A. baumannii cellular functions to identify potential targets for drug development has stalled due in part to laborious genetic techniques. Here we have pioneered a novel recombineering system that facilitates efficient genome editing in A. baumannii by single PCR products. This technology allows for rapid genome editing to quickly ascertain gene-phenotype relationships. To demonstrate the power of recombineering in dissecting A. baumannii biology, we use this system to establish key gene-phenotype relationships important to infection and persistence in hospitals, including oxidative stress protection, biocide resistance, and biofilm formation.

Dataset Information

Characterization of the chromosomal aac(6')-Ij gene of Acinetobacter sp. 13 and the aac(6')-Ih plasmid gene of Acinetobacter baumannii.

Publications

Characterization of the chromosomal aac(6')-Ij gene of Acinetobacter sp. 13 and the aac(6')-Ih plasmid gene of Acinetobacter baumannii.

Similar Datasets

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