TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Bacterial strain, plasmid, or primer | Description, relevant genotype, or sequencea |
|---|---|
| Bacterial strains | |
| B. melitensis | |
| 16M | Wild type |
| TAP1 (ΔvirB2) | ΔvirB2::kan |
| TAP4 (Δure1) | ΔureABC1::kan |
| TAP5 (Δpmm) | pmm::pNH1.3 |
| E. coli DH5α | Used for cloning |
| Plasmids | |
| pCR2.1-TOPO | TA cloning vector |
| pBluescript KS | Cloning vector |
| pUC4-KSAC | Plasmid with KSAC kanamycin resistance cassette |
| pUR | PCR products of ure fragments from nucleotides 1703818 to 1704817 and nucleotides 1707273 to 1708272 separated by KSAC in pBluescript KS |
| pAV2.2 | PCR products of virB fragments from nucleotides 988 to 1562 and nucleotides 1796 to 2458 separated by KSAC in pBluescript KS |
| pNH1.3 | PCR product of pmm nucleotides 355 to 610 in pBluescript KS |
| Primers | |
| BM1652upF | TCTAGAGGCTTGCAGGAGATTGAT (XbaI) |
| BM1652upR | GAATTCGCCGGAGTATGAGATATG (EcoRI) |
| BM1654dnF | GAATTCGGCGGCCATCGCGATCAA (EcoRI) |
| BM1654dnR | GTCGACCGTCAACGGGACCGGTGA (SalI) |
| PMM355F | GCTCCACCGAAACCGATGC |
| PMM610R | TCGCTTTTGCCCCATTGG |
a
The restriction sites underlined in the primer sequences are shown in parentheses after the sequences.