Highly conjugative pMG1-like plasmids carrying Tn1546-like transposons that encode vancomycin resistance in Enterococcus faecium - PubMed (original) (raw)

FIG. 1.

Agarose gel electrophoresis of restriction endonuclease-digested plasmid DNAs and hybridization with the pHTα probe. Southern hybridization was performed with the digoxigenin-based nonradioisotope system of Boehringer GmbH (Mannheim, Germany), and all procedures were based on the manufacturer's manual and standard protocols (34). (A1) Agarose gel electrophoresis of _Nde_I-digested plasmid DNAs isolated from vancomycin-resistant E. faecium or E. avium (VRE) isolates. (A2) The gel was Southern blotted and hybridized to pHTα. Lanes of panels A1 and A2: 1, _Hin_dIII-digested lambda DNA; 2, _Nde_I-digested pMG1; 3, _Nde_I-digested pHTα; 4 to 15, _Nde_I-digested plasmid DNAs from the strains FH1, FH2, FH3, FH4, FH5, FH6, FH7, FH8, FH9, FH10, FH11 and FH12, respectively. (B1) Agarose gel electrophoresis of _Nde_I-digested pHTα, pHTβ, and pHTγ plasmid DNA isolated from each transconjugant of FH1, FH4, and FH7, respectively. (B2) The gel was Southern blotted and hybridized to the pHTα probe. Lanes of panels B1 and B2: 1, _Hin_dIII-digested lambda DNA; 2, pMG1; 3, pHTα; 4, pHTβ; 5, pHTγ.