PMC

Restriction maps of pNV1.2 and pNVrpoB2. See text for details.

Alignment of the RIF-binding regions of RNAP β subunits among N. farcinica IFM 10152 (RpoB and RpoB2), N. asteroides IFM 0319^T (Nast), and M. tuberculosis H37Rv (Mtub). Amino acid substitutions are represented in reverse color. RIF-binding regions (clusters I and II) are boxed.

Construction of the ΔrpoB2 mutant. A. Strategy for making an in-frame, unmarked deletion of rpoB2. See text for details. Δ indicates a deletion allele. The ScaI half-site generated after ligation of a 4.9-kb ScaI-SphI fragment containing rpoB2 to pUC19 digested with HincII and SphI is shown in parentheses. B. Southern hybridization analysis of a ΔrpoB2 mutant. NotI-digested total DNAs extracted from the wild-type strain (lane 1) and a ΔrpoB2 mutant (lane 2) were probed with a 0.6-kb EcoRI fragment containing nfa46450 (short black bar in panel A).

Distribution of rpoB duplication among Nocardia strains. Total DNAs extracted from N. asteroides IFM 0319^T (lane 1), N. asteroides IFM 10159 (lane 2), N. asteroides IFM 10162 (lane 3), N. brasiliensis IFM 0236^T (lane 4), N. brasiliensis IFM 0406 (lane 5), N. brasiliensis IFM 10132 (lane 6), N. brasiliensis IFM 10160 (lane 7), N. farcinica IFM 0284^T (lane 8), N. farcinica IFM 10125 (lane 9), and N. farcinica IFM 10152 (lane 10) were digested with BamHI and probed with a 437-bp fragment containing the C-terminal region of rpoB. The probe was prepared from the total DNA of N. farcinica IFM 10152 by PCR using the primers NFrpoBF and NFrpoBR.