Tn4351 transposes in Bacteroides spp. and mediates the integration of plasmid R751 into the Bacteroides chromosome

Abstract

The gene for resistance to erythromycin and clindamycin, which is carried on the conjugative Bacteroides plasmid, pBF4, has been shown previously to be part of an element (Tn4351) that transposes in Escherichia coli. We have now introduced Tn4351 into Bacteroides uniformis 0061 on the following two suicide vectors: (i) the broad-host-range IncP plasmid R751 (R751::Tn4351) and (ii) pSS-2, a chimeric plasmid which contains 33 kilobases of pBF4 (including Tn4351) cloned into the IncQ plasmid RSF1010 and which is mobilized by R751. When E. coli HB101, carrying either R751::Tn4351 or R751 and pSS-2, was mated with B. uniformis under aerobic conditions, Emr transconjugants were detected at a frequency of 10-6 to 10-5 (R751::Tn4351) or 10-8 to 10-6 (R751 and pSS-2). In matings involving pSS-2, all Emr transconjugants contained simple insertions of Tn43S1 in the chromosome, whereas in matings involving R751::Tn4351, about half of the Emr transconjugants had R751 cointegrated with Tn4351 in the chromosome. Of the Emr transconjugants, 13% were auxotrophs.Bacteroides spp. which had R751 cointegrated with Tn4351 in the chromosome did not transfer R751 or Tn4351 to E. coli HB101 or to isogenic B. uniformis, nor did the integrated R751 mobilize pE5-2, an E. coli-Bacteroides shuttle vector that contains a transfer origin that is recognized by R751.

Changes of resistome, mobilome and potential hosts of antibiotic resistance genes during the transformation of anaerobic digestion from mesophilic to thermophilic

“…As described previously (Tian et al, 2015), Bacteroides was the dominant fermentative bacteria in mesophilic sludge, and experienced fast decline after temperature increase because all species of this genus are mesophilic (Garrity et al, 2004). At the same time, tet(X) has not yet been proved to be horizontally transferred to other strains (Ghosh et al, 2009b), in spite of the fact that it has been linked with a variety of mobile genetic elements (Guiney et al, 1984;Robillard et al, 1985;Shoemaker et al, 1986;Ghosh et al, 2009b). Therefore, the high reduction efficiency of tet(X) during thermophilic AD could be attributed to the elimination of its bacterial host by the high temperature.…”

Comparison of Bacteroides thetaiotaomicron and Escherichia coli 16S rRNA gene expression signals

“…All of the E. coli-Bacteroides lux reporter/ shuttle vectors were transferred into BT4001 by diparental mating via E. coli strain S17-1 (Simon et al, 1983) using standard filter-mating protocols (Shoemaker et al, 1986).…”

Characterizing the Interactions between a Naturally Primed Immunoglobulin A and Its Conserved Bacteroides thetaiotaomicron Species-specific Epitope in Gnotobiotic Mice

“…Whole Genome Transposon Mutagenesis-Mutagenesis of B. thetaiotaomicron VPI-5482 was performed using pEP4351 (pir-dependent R6K oriV; RP4 oriT; Cm r Tc r (Em r ); Tn4351 mutagenesis vector (31,32)). Mutants were isolated based on their growth on BHI blood agar plates containing erythromycin (10 g/ml to select for transposon-containing B. thetaiotaomicron strains) and 600 g/ml gentamicin (to select against any persistent E. coli harboring the transposon mutagenesis vector).…”