Physical characterisation of the replication region of the Streptococcus faecalis plasmid pAMβ1

Swinfield, Tracy-Jane; Oultram, John D.; Thompson, D.E.; Brehm, John K.; Minton, Nigel P.

doi:10.1016/s0378-1119(19)30488-3

Cited by 116 publications

(66 citation statements)

References 33 publications

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“…Plasmids were transferred from gels to nylon filters by a vacuum procedure, followed by hybridization with the hyl Efm , as described elsewhere [19]. Matings designed to transfer plasmid pAM␤1 into GE-1 were performed using E. faecalis OG1X (pAM␤1) [20] as a donor, as above, with selection on BHI agar plates containing rifampin, fusidic acid, and erythromycin (10 g/mL). Mouse gastrointestinal colonization model.…”

Section: Methodsmentioning

confidence: 99%

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Transferable Capacity for Gastrointestinal Colonization inEnterococcus faeciumin a Mouse Model

et al. 2009

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A high level of gastrointestinal colonization frequently precedes invasive infection due to Enterococcus faecium. Factors other than antimicrobial resistance that promote gastrointestinal colonization by E. faecium have not been identified. We tested the ability of a colonization-proficient clinical E. faecium isolate (C68) to transfer colonizing ability to noncolonizing E. faecium recipient strains. Transconjugants derived from matings that used E. faecium D344SRF as a recipient strain colonized mouse gastrointestinal tracts in high numbers under selective pressure from clindamycin or vancomycin, compared with control strains that lacked DNA transferred from C68. We transferred DNA into a second recipient strain (E. faecium GE-1), which also colonized mice in significantly greater numbers under selective pressure from clindamycin, compared with a control strain. These results indicate that E. faecium clinical isolates express transmissible factors other than antimicrobial resistance that promote colonization of the mouse gastrointestinal tract.

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Section: Methodsmentioning

confidence: 99%

“…To control for the expression of erythromycin resistance in these experiments, we mated GE-1 with E. faecalis OG1X (pAM␤1), with selection for transfer of erythromycin resistance, which is conferred by the ermB located on pAM␤1 [20]. One transconjugant was selected and designated GE-1B1.…”

Section: Figurementioning

confidence: 99%

Transferable Capacity for Gastrointestinal Colonization inEnterococcus faeciumin a Mouse Model

et al. 2009

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“…The right end of Tn4001 within Tn5385 interrupts sequences identical to the replication region (Rep in fig. 1) of broad host-range enterococcal plasmid pAMb1 [29], which itself has been interrupted by a deletion derivative of erythromycin resistance transposon Tn917. This replication region is likely nonfunctional, since the region known to be the origin of pAMb1 replication has been deleted by the insertion of IS256 [29].…”

Section: Tn5385mentioning

confidence: 99%

“…1) of broad host-range enterococcal plasmid pAMb1 [29], which itself has been interrupted by a deletion derivative of erythromycin resistance transposon Tn917. This replication region is likely nonfunctional, since the region known to be the origin of pAMb1 replication has been deleted by the insertion of IS256 [29]. To the right of the replication region gene repC lie directly repeated copies of staphylococcal IS element IS257 (identical to IS431 -see above) flanking a mercury resistance operon (Mer in fig.…”

Section: Tn5385mentioning

confidence: 99%

Association of different mobile elements to generate novel integrative elements

Rice¹

2002

Cellular and Molecular Life Sciences (CMLS)

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Among the more important problems in modern hospitals is the prevalence of bacterial pathogens expressing resistance to multiple antimicrobial agents. The frequency of multiresistance suggests mechanisms by which bacterial species can concentrate and efficiently exchange a variety of resistance determinants. Mechanisms by which this occurs include insertion of transposons within transposons, coalescence through the activity of insertion sequences and the employment of integrons. In some instances, more than one of these mechanisms is involved in creating large multiresistance genetic elements. The association of the elements with transferable elements or transposons may promote rapid dissemination among clinical strains, and create further opportunities for inclusion of additional resistance determinants.

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“…Although limited by the inability of certain cloned DNA sequences to generate co-integrates with conjugarive plasmids, the fact that these plasmids are stably replicated, in contrast to those that replicate by the rolling circle method (Swinfield et al 1990), makes them an attractive vehicle for the construction of recombinant molecules in lactobacilli. Problems associated with cointegrate generation may well be overcome when the molecular mechanisms at work are fully understood.…”

Section: Expression Of the Fl-glucanase Genementioning

confidence: 99%

Molecular cloning in Lactobacillus helveticus by plasmid pSA3::pVA797 co-integrate formation and conjugal transfer

Thompson¹,

Collins

1991

Appl Microbiol Biotechnol

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A gene encoding beta-glucanase activity from Bacillus amyloliquefaciens was subcloned in both orientations into plasmid shuttle vector pSA3. In only one orientation could a co-integrate be generated with the conjugative plasmid pVA797. The plasmid co-integrate was conjugated into Lactobacillus helveticus strain CNRZ450, where it was stably maintained without antibiotic selection and exhibited beta-glucanase activity. This method of introducing cloned DNA into thermophilic lactobacilli will facilitate the study of heterologous gene expression in non-transformable species.

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Physical characterisation of the replication region of the Streptococcus faecalis plasmid pAMβ1

Cited by 116 publications

References 33 publications

Transferable Capacity for Gastrointestinal Colonization inEnterococcus faeciumin a Mouse Model

Transferable Capacity for Gastrointestinal Colonization inEnterococcus faeciumin a Mouse Model

Association of different mobile elements to generate novel integrative elements

Molecular cloning in Lactobacillus helveticus by plasmid pSA3::pVA797 co-integrate formation and conjugal transfer

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