Sequential Isolation in a Patient of Raoultella planticola and Escherichia coli Bearing a Novel ISCR1 Element Carrying blaNDM-1
Figure 2
The characterization and mobilization model of the 4812-1-ISCR 1 element.
(a): Genetic context of plasmids pEcNDM1-4 and pEcNDMneg-4 and mobilization of the 4812 bp NDM-1-ISCR1 element. Red boxes denote 4812 bp NDM-1-ISCR1 element; Horizontal arrow denote the direction of primers; Red downward pointing arrows denote NotI restriction site; Blue upward pointing arrows denote ClaI restriction site; Purple upward pointing arrows BsaX1 restriction site; Lowercase red letters denote corresponding fragments in (f) and (g). (b): PFGE of Xba I digested patterns of E. coli strains EcNDM1 and EcNDMneg. (c): PFGE of S1 digested plasmid profiles from E. coli EcNDM1, R. planticola RpNDM1 and transformant E. coli JM109 (pEcNDM1-4). (d): Southern blot hybridization of (c) with a blaNDM-1 probe. (e): PFGE of plasmid pEcNDM1-4 with or without restriction enzyme digestion stained with ethidium bromide. (f): Southern blot hybridization of (e) with a blaNDM-1 probe. Lowercase red letters denote fragments expected as depicted in (a). (g): PCR amplification for confirmation of the structure of the NDM-1-ISCR1 element with primer pairs as shown in (a).