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. 2010 Oct 15;192(24):6346–6351. doi: 10.1128/JB.00838-10

FIG. 2.

FIG. 2.

Excision of GI-2 occurs spontaneously. (A) Long-range PCR analysis (primers P1 and P2) (Fig. 1) for the presence of GI-2 in Brucella cultures. Bo, B. ovis; Ba, B. abortus 2308; Bm, B. melitensis 16M; Bs, B. suis 1330. Abbreviations: 17.3-kb GI-2, GI-2-containing fragment (Fig. 1); 2.2-kb ΔGI-2, chromosomal scar carrying fragment (Fig. 1); M, λ/HindIII DNA ladder (Fermentas). (B) PvuII restriction analysis of the 2.2-kb fragment of the following: B10, B. abortus B10, and Bo, B. ovis. M, 1-kb DNA ladder (Fermentas). (C) PCR analysis of R B. abortus mutants R1 and R6. (Top) Detection of the chromosomal scar with primers P5 and P6 (Fig. 1); (bottom) detection of the int gene with primers P3 and P4 (Fig. 1). M, 100-bp DNA ladder (Promega, Madison, WI). (D) IS711 Southern blot of the following: B10, B. abortus B10 (field isolate); R6, R B. abortus mutant derived from B10; Ba, B. abortus 2308; and R1, R B. abortus mutant derived from strain 2308. The arrow shows the position of the 4.2-kb ClaI restriction fragment (Fig. 1) that carries the IS711 copy. M, λ/HindIII biotin-labeled DNA ladder. All experiments were performed with DNA from broth cultures, except for 2C, in which DNA from a single bacterial colony was used.