FIG 2.

Role of 84 in MGIVchHai6 mobilization. (A) Mobilization of MGI^Kn or its Δ84 mutant by pVCR94^Sp. (B) Schematic representation of the 85-84 region of MGIVchHai6. Genes are color coded as indicated in Fig. 1A. The position and sequence of alternative start codons within 84 are indicated. (C) Complementation assays of the Δ84 mutation by alternative ORFs within 84. When indicated, donor strains contained the complementation plasmid pBAD-84 or one of its derivatives (Table 1). (D) Confirmation of ATG₁₁₈ as the genuine start codon of mobI_M. Conjugation assays were performed using E. coli GG56 (Nx) containing the specified elements as donor strains and either CAG18439 (Tc) (A and C) or VB112 (Rf) (D) as the recipient strain. The bars represent the means ± standard errors of the means from three independent experiments. Statistical analyses were carried out on the logarithm of the values using a one-way ANOVA followed by Dunnett’s multiple-comparison test with the WT MGI^Kn (A), pBAD-84 (C), or pIG0-84 (D) as the control. Statistical significance is indicated as follows: ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; ns, not significant.