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. 2009 Jun 29;53(9):3675–3682. doi: 10.1128/AAC.00550-09

FIG. 1.

FIG. 1.

Southern blot analysis for p55 gene inactivation. Genomic DNA isolated from the M. bovis BCG wild-type (WT), M. bovis BCG single-crossover (SXO), and M. bovis BCG double-crossover (DXO) strains was digested with PvuII and hybridized to a probe corresponding to a 0.9-kb p55 internal region. The DXO strain carrying the inactivated p55 gene lost the wild-type 5-kb band and gained a 7.3-kb band, reflecting the insertion of the 2.3-kb hygromycin resistance cassette (cf. lane WT and lane DXO). The SXO strain showed two bands (9.3 and 7.2 kb) due to the incorporation of the suicide vector in the genome. The diagram showing the expected PvuII digestion fragment is not shown to scale. Ωhyg, cassette containing the hygromycin resistance gene; pGOAL17, cassette containing the lacZ and sacB genes; Km, kanamycin resistance gene from p2NIL vector.