Table 1. Plasmid-borne rpoB482 variants from different Bacillus species or subspecies cloned into E. coli pUC57 vector.
The house-keeping rpoB gene encodes the essential β subunit of RNA polymerase. A single C to T transition at codon 482 (position 1446) in the rpoB gene confers resistance to rifampicin (RifR) (Nicholson and Maughan, 2002). This mutation was introduced in the rpoB region of the indicated Bacillus species or subspecies leading to the rpoB482 mutant variants for selection of the RifR transformants (see Figure 1A). Homologous regions at 5′- and 3′-ends of the different rpoB482 genes were selected to eliminate variants in the PCR amplification efficiencies. The 2,997-bp segment of the different rpoB482 genes was in vitro synthesized and cloned into Escherichia coli 2,710-bp pUC57 vector (Ampicillin resistance). The accuracy of the in vitro synthesized DNAs was confirmed by nucleotide sequence (GeneWiz, London, UK).
| Plasmid name | Origin of Bacillus DNAs | Number of mismatches |
Donor-recipient sequence divergence (in %) |
| pCB980 | B. subtilis 168 | 1 | 0.03 |
| pCB981 | B. subtilis W23 | 74 | 2.47 |
| pCB982 | B. atrophaeus 1942 | 250 | 8.35 |
| pCB983 | B. amyloliquefaciens DSM7 | 303 | 10.12 |
| pCB984 | B. licheniformis DSM13 | 435 | 14.52 |
| pCB1054 | B. gobiensis FJAT4402 | 510 | 17.0 |
| pCB985 | B. thuringiensis MC28 | 624 | 20.83 |
| pCB1056 | B. smithii DSM4216 | 681 | 22.74 |