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. 1999 Dec;181(23):7314–7322. doi: 10.1128/jb.181.23.7314-7322.1999

FIG. 2.

FIG. 2

Confirmation of rocF allelic exchange mutants of H. pylori. (A) Southern blot analysis of H. pylori WT and the rocF::aphA3 isogenic mutant. Chromosomal DNA (∼6 μg) from H. pylori 26695 and 26695 rocF::aphA3 was digested with AvaI and processed as described in Materials and Methods. The blot was probed with either the kanamycin cassette (1.2 kb) or a portion of the rocF gene (240-bp HindIII fragment). Note that when probed with rocF, the mutant has an increase in molecular weight by ∼1.2 kb, as expected. (B) PCR analysis of rocF mutants of H. pylori. The primer pair strategies used are shown in Table 2. Lanes (number, H. pylori strain, primer pair): 1, N6 WT, R13 and K4; 2, N6 rocF::aphA3, R13 and K4; 3, N6 236-2, R13 and K4; 4, SS1 WT, R13 and K4; 5, SS1 rocF::aphA3, R13 and K4; 6, SS1 236-2, R13 and K4; 7, N6 WT, R5 and K8; 8, N6 rocF::aphA3, R5 and K8; 9, N6 WT, R17 and K8; 10, N6 236-2, R17 and K8; 11, SS1 WT, R17 and K5; 12, SS1 rocF::aphA3, R17 and K5; 13, SS1 WT, R17 and K8; 14, SS1 236-2, R17 and K8; and M, 1-kb marker (Gibco BRL).