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A total of seven different ClaI-arcCB DNA restriction band patterns were identified among the 39 S. epidermidis strains carrying ACME (Table 1).
In order to determine BAC order and orientation, we characterized all BACs in the libraries using high resolution restriction band fingerprinting.
ALAD alleles are differentiated based on the existence of a MSPI endonuclease restriction site specific to the ALAD-2 derived ALAD-2 derivede chain reaction) fragment, which yields a diagnostic restriction band.
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LNT01 (recipient), wild type and plasmid-tagged donor strains, and three clones of each transconjugant all exhibited unique restriction banding patterns when digested with SmaI, XhoI or NarI.
The KpnI restriction bands of 5.0 and 6.3 kb present in P1-mm and absent from P1-wr are derived from the two unique P1-mm gene copies.
PFGE analysis of digested samples allowed us to use the unique restriction banding patterns of the strains as a genetic screen to visually determine whether the plasmids were transferred to the recipient strains.
The DNA restriction banding pattern of the wild type strain LNT01 (lane 7A) digested with SmaI was identical to that of the transconjugants (lanes 10 12A), except that the banding pattern of all transconjugant clones contained an additional band of ∼270 kb.
The abundance distribution of the restriction bands in all these BAC fingerprints is presented in Figure 1.
Accessions with no restriction bands on a LI-COR gel were assigned to the reference haplotype H0 (93161P).
On the other hand, restriction of the two kanamycin resistant clones yielded fully overlapping restriction bands, indicating identity of the inserts.
On average, each BAC clone contained 98.7 restriction bands, with a range of 60 to 120 bands from most of the BACs.
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