Article Figures & Data
Figures
- Fig. 1.
Analysis of genomic DNA from E. faeciumHM1025 and HM1025-1, digested with SmaI (A and B, left) and from E. faecalis JH2-2 and a transconjugant digested withSfiI (A and B, right) by pulsed-field gel electrophoresis and hybridization. (A) Lanes 1, E. faecium HM1025; lanes 2,E. faecium HM1025-1. (B) Lanes 1, E. faecalisJH2-2; lanes 2, E. faecalis JH2-2/L1 (transconjugant resistant to erythromycin, lincomycin, gentamicin, streptomycin, and tetracycline). The digested fragments were transferred to a nylon sheet and hybridized to an in vitro digoxigenin-labeled linBprobe. Numbers on the left of the gels are molecular sizes in kilobases.
- Fig. 2.
Mechanism whereby clindamycin and lincomycin are converted to lincomycin and clindamycin 3-(5′-adenylate) by LinB in the presence of ATP and MgCl2+.
- Fig. 3.
Autoradiogram ofl-[35S]methionine-labeled polypeptide specified in vitro by the amplification product of linB. (A) Protein electrophoresis in a 15% polyacrylamide gel containing sodium dodecyl sulfate. After separation, proteins were stained with Coomassie blue. (B) Visualization of labeled protein bands in the gel by autoradiography. Lanes 1, control without template; lanes 2, protein products synthesized from PCR-generated linB DNA; lanes 3, molecular mass markers (masses on the left are expressed in kilodaltons). The position of LinB (31,195 Da) is indicated.
Tables
- Table 1.
MICs of erythromycin and lincosamides against bacterial strains
Organism MICs (μg/ml) of: Origin of strain or reference Erythromycin Lincomycin Clindamycin E. faeciumHM1025 >128 >128 >128 This study E. faeciumHM1025-1 0.25 0.25 0.12 Cured derivative of E. faecium HM1025 E. faecalisJH2-2 0.12 8 2 Recipient strain E. faecalisJH2-2/1025 >128 >128 >128 Transconjugant from E. faecium HM1025 E. coliDB10 4 8 0.25 Datta et al. (9) E. coli DB10/pVMM25 4 64 8 This study E. coli DB10/pVMM27 4 64 8 This study S. aureus RN4220/pJIM2246 0.25 0.5 0.06 This study S. aureus RN4220/pVMM26 0.25 64 0.5 This study
