TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristic(s)a | Source or reference |
|---|---|---|
| B. abortus strains | ||
| 2308 | Wild type | B. Deyoe |
| BA1143 | virB4::lacZYA | This work |
| ADH1 | ΔvirB2::Km (polar) | This work |
| ADH3 | ΔvirB2 (nonpolar) | This work |
| ADH4 | ΔvirB1-12::Km | This work |
| ADH5 | ΔvirB1::Km (polar) | This work |
| ADH6 | ΔvirB1 (nonpolar) | This work |
| ADH8 | Knock-in complementation of ADH3 | This work |
| E. coli strains | ||
| DH5α | endA 1 hsdR17 (rκ− mκ−) supE44 thi-1 recA1 gyrA relA1 Δ(lacZYA-argF)U169 deoR (φ80 dlac Δ[lacZ]M15) | 34 |
| BL21 (DE3) | F′ompT gal [dcm] [lon] hsdSB (rB− mB−) DE3 | 31 |
| Plasmids | ||
| pBluescript KS | ColE1, bla | Stratagene |
| pCR2.1-TOPO | TA cloning vector | Invitrogen |
| pUC4-KSAC | Carries Tn903 kanamycin resistance cassette | Pharmacia |
| pAV1.4 | virB219-477 and virB1154-1562 cloned into pBluescript | This work |
| pAV1.5 | virB219-477 and virB1154-1562 separated by kanamycin resistance cassette KSAC cloned into pBluescript | This work |
| pAV2.1 | PCR products of virB988-1562 and virB1796-2458 cloned into pBluescript | This work |
| pAV2.2 | PCR products of virB988-1562 and virB1796-2458 separated by kanamycin resistance cassette cloned into pBluescript | This work |
| pAS1.1 | sacB cloned into pAV2.1 | This work |
| pAS1.2 | sacB cloned into pAV1.4 | This work |
| pMR10 | RK2, orivV + trfA, oriT, nptI, lacZα | R. M. Roop II |
| pEX18Ap | sacB | 17 |
| pHB11 | virB140-1967 cloned into pMR10 | This work |
| pFUSE | oriR6K plasmid containing promoterless lacZYA | 5 |
| pBA1143 | virB1644-2457 cloned into pFUSE | This work |
| pAVT1.4 | PCR products of B. abortus using primer pairs 98736F-99854R and 112902F-114022R separated by kanamycin resistance cassette cloned into pBluescript | This work |