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. 2010 Sep;156(Pt 9):2829–2841. doi: 10.1099/mic.0.040675-0

Fig. 1.

Fig. 1.

Construction of an M. tuberculosis ΔpknK strain by allelic replacement. (a) Schematic representation of the chromosomal locus of pknK and its adjacent genes in the wild-type H37Rv and the LIX11 mutant strain, in which the pknK gene is deleted and replaced with a hygromycin-resistance cassette. The striped bars indicate the position of the DNA probes used for Southern blot analysis and the dashed lines indicate the expected size of the hybridization with the specific probes. (b) Southern blot confirmation of the LIX11 mutant strain. Chromosomal DNA was isolated from strains H37Rv (lane 1) and LIX11 (lane 2), digested with HindIII and hybridized with probes specific for the hyg, pknK and 3′ flank regions. (c) Western blot analysis of PknK (∼119 kDa, arrow) in the whole-cell lysates of M. tuberculosis H37Rv (lane 1), LIX11 mutant (lane 2) and LIX16 complemented mutant (lane 3) strains. M, molecular size markers.