Characterization of the Acinetobacter Plasmid, pRAY, and the Identification of Regulatory Sequences Upstream of an aadB Gene Cassette on This Plasmid

Segal, Heidi; Bg, Elisha

doi:10.1006/plas.1999.1403

Cited by 26 publications

(27 citation statements)

References 24 publications

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“…The aadB gene (conferring resistance to tobramycin) in AB058 is on the plasmid pRAY (35) and is not associated with an IS element or transposon. This is in contrast to the location on AbaR1 in AYE.…”

Section: Resultsmentioning

confidence: 99%

Genomewide Analysis of Divergence of Antibiotic Resistance Determinants in Closely Related Isolates of Acinetobacter baumannii

Adams

Chan²,

Molyneaux³

et al. 2010

Antimicrob Agents Chemother

101

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Multidrug resistance has emerged as a significant concern with infections caused by Acinetobacter baumannii. Ample evidence supports the involvement of mobile genetic elements in the transfer of antibiotic resistance genes, but the extent of variability and the rate of genetic change associated with the acquisition of antibiotic resistance have not been studied in detail. Whole-genome sequence analysis of six closely related clinical isolates of A. baumannii, including four from the same hospital, revealed extensive divergence of the resistance genotype that correlated with observed differences in antimicrobial susceptibility. Resistance genes associated with insertion sequences, plasmids, and a chromosomal resistance gene island all showed variability. The highly dynamic resistance gene repertoire suggests rapid evolution of drug resistance.

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Section: Resultsmentioning

confidence: 99%

Genomewide Analysis of Divergence of Antibiotic Resistance Determinants in Closely Related Isolates of Acinetobacter baumannii

Adams

Chan²,

Molyneaux³

et al. 2010

Antimicrob Agents Chemother

101

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“…Integrons show a strong preference for capture of gene cassettes by recombination at their cognate attI site (Collis et al, 2002b), although cassette capture by recombination at 59-be sites or secondary sites (not associated with integrons or gene cassettes) is known (Francia et al, 1993;Recchia et al, 1994) and expression from promoters upstream of secondary sites has been demonstrated (Segal & Elisha, 1999). We screened three of the unknown-Gm R strains (Q23-10, Q23-12 and Q23-13) for insertion of pUS23 at the 59-be sites associated with the 10 gene cassettes in the InPstQ array by PCR (data not shown), but the results indicated that pUS23 had not integrated into the known cassette array.…”

Section: Discussion the Significance Of Cismentioning

confidence: 99%

The native Pseudomonas stutzeri strain Q chromosomal integron can capture and express cassette-associated genes

Coleman

Holmes

2005

Microbiology

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The integron-gene cassette system contributes to multiple antibiotic resistance in bacteria and is likely to be of broader evolutionary significance. However, the majority of integron diversity consists of chromosomal integrons (CIs), with mostly unknown phenotypes, which are poorly characterized. A pUC-based reporter plasmid (pUS23) was developed containing a recombination site [aadB 59 base element (59-be)] upstream of promoterless aadB [gentamicin (Gm) resistance] and gfp (green fluorescence) genes, and this construct was used to investigate the recombination and expression activities of the CI in Pseudomonas stutzeri strain Q. Electroporation of pUS23 into P. stutzeri Q gave ampicillin-resistant transformants, which yielded Gm R green fluorescent recombinants after plating on Gm medium. Site-specific integration of pUS23 at attI was detected by PCR in 8 % of Gm R colonies and the frequency of attI integration was estimated as 2?0610 "8 per P. stutzeri Q(pUS23) cell. RT-PCR confirmed integron-mediated expression of aadB in one recombinant strain (Q23-17) and a promoter (P c ) was localized to the 59 end of the intI gene. The integrated pUS23 and flanking integron DNA were cloned from genomic DNA of strain Q23-17 and sequenced, confirming that site-specific integration of the entire reporter plasmid had occurred at the attI site. An insertion sequence (ISPst5; IS5 family) was discovered in the vector backbone of the reporter plasmid integrated at attI and also in a pUS23 derivative recovered as a plasmid in Escherichia coli JM109. This is the first demonstration that wild-type CIs can capture gene cassettes and express cassette-associated genes. INTRODUCTIONIntegrons and gene cassettes function together as a distinctive genetic system (Fig. 1). Gene cassettes typically consist of a single promoterless gene and an associated recombination site (termed 59 base element, 59-be or attC).The total pool of cassette-associated genes in natural bacterial populations (gene cassette metagenome) is a significant source of genetic novelty (Holmes et al., 2003a;Michael et al., 2004;Stokes et al., 2001), and since individually packaged genes create the possibility of combinatorial genetics, the significance of the gene cassette metagenome is potentially far reaching. However, gene cassettes are dependent on externally supplied functions for both mobilization and expression. These functions are supplied by integrons Rowe-Magnus & Mazel, 2001). The key features of an integron are a recombination site (attI), an enzyme (integron integrase, IntI) that catalyses site-specific recombination between cassette and integron sites (attI659-be or 59-be659-be) and a cassette promoter (P c ). Collectively, these give an integron the capacity to acquire gene cassettes, express genecassette-associated ORFs (gcORFs) and rearrange the transcriptional order of gcORFs through repeated integration and excision of gene cassettes. evidence is hard to find, suggesting that other integron classes have been less successful in this regard than class 1...

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“…Likewise, Fig. 2 shows the coverage of the antibiotic resistance plasmid pRAY originating from the clinical isolate Acinetobacter strain SUN (Segal and Elisha, 1999). The pRAY module containing mobilisation genes and three orfs of unknown function is covered by more plasmid metagenome reads than the rest of the plasmid indicating higher abundance of the corresponding module in the sequenced sample.…”

Section: Mapping Of Wwtp Plasmid Metagenome Reads To Plasmid Genes Anmentioning

confidence: 98%

“…Other abundant genes are very similar to the partitioning gene parA of the mobilisable IncP-6 plasmid Rms149 of Pseudomonas aeruginosa (Haines et al, 2005), the replication initiation gene of the Salmonella enterica subsp. enterica colicigenic plasmid pBERT (NC 001848), the replication gene rom of the Klebsiella pneumoniae bacteriocinproducing plasmid pKlebB-K17/80 (Riley et al, 2001) and a gene with unknown function of the antibiotic resistance plasmid pRAY of a clinical Acinetobacter isolate (Segal and Elisha, 1999). Further prominent sequence reads originate from transposable elements such as the chromate resistance transposon Tn5719 (Tauch et al, 2003), the tetracycline resistance transposon Tn1721 (Allmeier et al, 1992;Rhodes et al, 2004), the transposon Tn5403 of the extended-spectrum ␤-lactamase encoding plasmid pC15-1a (Boyd et al, 2004), Tn3-related transposons (Rhodes et al, 2004) and an insertion sequence of the IS5 family.…”

Section: Annotation Of Single Reads Of the Wwtp Plasmid Metagenomementioning

confidence: 99%

Insight into the plasmid metagenome of wastewater treatment plant bacteria showing reduced susceptibility to antimicrobial drugs analysed by the 454-pyrosequencing technology

Szczepanowski

Bekel

Goesmann

et al. 2008

Journal of Biotechnology

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Characterization of the Acinetobacter Plasmid, pRAY, and the Identification of Regulatory Sequences Upstream of an aadB Gene Cassette on This Plasmid

Cited by 26 publications

References 24 publications

Genomewide Analysis of Divergence of Antibiotic Resistance Determinants in Closely Related Isolates of Acinetobacter baumannii

Genomewide Analysis of Divergence of Antibiotic Resistance Determinants in Closely Related Isolates of Acinetobacter baumannii

The native Pseudomonas stutzeri strain Q chromosomal integron can capture and express cassette-associated genes

Insight into the plasmid metagenome of wastewater treatment plant bacteria showing reduced susceptibility to antimicrobial drugs analysed by the 454-pyrosequencing technology

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