Conjugative R plasmids in Streptococcus agalactiae (group B)

Horodniceanu, T; Bougueleret, Lydie; El-Solh, Névine; Bouanchaud, D. H.; Chabbert, Y

doi:10.1016/0147-619x(79)90038-6

Cited by 77 publications

(64 citation statements)

References 17 publications

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“…The inducible chloramphenicol resistance determinant from the 67-kb conjugative ⍀ element of Streptococcus agalactiae B109 was provided as a 2.3-kb HindIII fragment cloned into pBR322 and provided resistance in both S. agalactiae and E. coli (18,22). To further reduce the size of the insert, a Sau3A partial digest of this plasmid was performed and the resulting fragments were cloned into the BamHI site of pBluescript SK ϩ .…”

Section: Resultsmentioning

confidence: 99%

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Analysis of promoters in Borrelia burgdorferi by use of a transiently expressed reporter gene

1997

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A transient chloramphenicol acetyltransferase (CAT) expression system was developed for Borrelia burgdorferi. An Escherichia coli vector containing a promoterless Streptococcus agalactiae cat gene was constructed. Promoters for ospA, ospC, and flaB were placed upstream of this cat gene, and CAT assays were performed in E. coli from these stably maintained plasmids. The plasmids with putative promoters ospA and flaB were found to be approximately 20-fold more active than were the plasmids with ospC or no promoter. The level of activity correlated well with the resistance to chloramphenicol that each plasmid provided. Next, the nonreplicative plasmid constructs were transformed by electroporation into B. burgdorferi. CAT assays were performed by both thin-layer chromatography and the fluor diffusion method. Measurement of CAT activity demonstrated that the ospA promoter was again about 20-fold more active than the promoterless cat gene. The flaB and ospC promoters increased the activity seven-and threefold, respectively, over that with the promoterless construct. This simple transient-expression assay was shown to be an effective method to study promoter function in B. burgdorferi in the absence of a well-developed genetic system.

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Section: Resultsmentioning

confidence: 99%

“…Construction of plasmids. The source of the cat gene was Streptococcus agalactiae B109 (18,22), kindly provided by J. Inamine, Colorado State University, Fort Collins, Colo. Partial digests of this cat gene were performed by published methods (26).…”

Section: Methodsmentioning

confidence: 99%

Analysis of promoters in Borrelia burgdorferi by use of a transiently expressed reporter gene

1997

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“…BM5320 (7), BM5324 (7), and BM5329 (this study) are Cmr Emr Kmr Tcr_Mnr transconjugants obtained from the mating of BM4200 with E. faecalis, S. pneumoniae, and S. sanguis recipients, respectively. Twenty-three clinical isolates of streptococci of groups A, B, C, and G as well as three strains, BM6502 (strain 14) (3), BM6103 (strain 16) (29), and BM6106 (strain 17) (29), which were obtained by curing their R plasmids from the original wild-type strains C87 (3), B96 (25), and B97 (27), respectively, are presented in Table 3. Five other streptococci were also studied: four of group A (A453 [28], A456 [6], A472 [6], and BM6402 [5]) and one of group B (B126 [41] (25,27).…”

Section: Methodsmentioning

confidence: 99%

“…The streptococcal strains used in this study were antibiotic-resistant clinical isolates. Four of these strains carry conjugative chromosomal elements: S. pyogenes A454 (26,34) contains Tn3701 (33) encoding Emr Tcr-Mnr, S. agalactiae B109 (26,27) contains Tn3951 (31) encoding Cmr Em' Tc'-Mn', S. pneumoniae BM6001 (7,15,45) contains the element Ql(cat-tet) (48) encoding Cmr Tcr_Mnr, and S. pneumoniae BM4200 (7) contains Tn1545 (9) encoding Emr Kmr Tcr-Mnr. BM4200 is also resistant to chloramphenicol, penicillins, and trimethoprim (7,41).…”

Section: Methodsmentioning

confidence: 99%

Presence of chromosomal elements resembling the composite structure Tn3701 in streptococci

1990

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Tn3701, carried by Streptococcus pyogenes A454, is the first chromosomal composite element to be described; it contains in its central region Tn3703, a transposon similar to Tn916. A comparison by DNA-DNA hybridization of Tn3701 with omega(cat-tet) and Tn3951, carried by Streptococcus pneumoniae BM6001 and by Streptococcus agalactiae B109, respectively, revealed that the two latter structures are also Tn3701-like composite elements. The DNAs of 27 other antibiotic-resistant group A, B, C, and G streptococci and of S. pneumoniae BM4200 showed sequence homologies to Tn3701 (14 strains, including BM4200), to the regions of Tn3701 outside of Tn3703 (5 strains), and to Tn916 alone (8 strains). The DNAs of five strains did not detectably hybridize with any probe. The tetM gene was identified in most chromosomal genetic elements coding for tetracycline-minocycline resistance. Since Tn3701-like elements are widely disseminated among antibiotic-resistant streptococci (47% of the 34 strains studied), we propose that Tn3701 be considered the prototype of chromosomal composite elements.

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“…pIP501, a multiple antibiotic resistance plasmid, was originally isolated from Streptococcus agalactiae (Horodniceanu et al, 1979). It has the broadest known host range for plasmid transfer in Grampositive bacteria and is furthermore the first conjugative plasmid originating from Gram-positive bacteria for which stable replication in Gram-negative bacteria has been shown (Kurenbach et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Crystallization and preliminary structure determination of the transfer protein TraM from the Gram-positive conjugative plasmid pIP501

et al. 2013

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The major means of horizontal gene spread (e.g. of antibiotic resistance) is conjugative plasmid transfer. It presents a serious threat especially for hospitalized and immuno-suppressed patients, as it can lead to the accelerated spread of bacteria with multiple antibiotic resistances. Detailed information about the process is available only for bacteria of Gram-negative (GÀ) origin and little is known about the corresponding mechanisms in Gram-positive (G+) bacteria. Here we present the purification, biophysical characterization, crystallization and preliminary structure determination of the TraM C-terminal domain (TraMÁ, comprising residues 190-322 of the full-length protein), a putative transfer protein from the G+ conjugative model plasmid pIP501. The crystals diffracted to 2.5 Å resolution and belonged to space group P1, with unitcell parameters a = 39.21, b = 54.98, c = 93.47 Å , = 89.91, = 86.44, = 78.63 and six molecules per asymmetric unit. The preliminary structure was solved by selenomethionine single-wavelength anomalous diffraction.

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Conjugative R plasmids in Streptococcus agalactiae (group B)

Cited by 77 publications

References 17 publications

Analysis of promoters in Borrelia burgdorferi by use of a transiently expressed reporter gene

Analysis of promoters in Borrelia burgdorferi by use of a transiently expressed reporter gene

Presence of chromosomal elements resembling the composite structure Tn3701 in streptococci

Crystallization and preliminary structure determination of the transfer protein TraM from the Gram-positive conjugative plasmid pIP501

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