TABLE 1.
Bacterial strains and plasmidsa
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Brucella abortus | ||
| 2308 | Wild type | B. Deyoe |
| ADH4.2 | ΔvirB1-virB12 | 19 |
| ADH6 | ΔvirB1 (nonpolar) | 19 |
| ADH3 | ΔvirB2 (nonpolar) | 19 |
| ADH18 | ΔvirB3::Km (polar) | This work |
| ADH19 | ΔvirB3 (nonpolar) | This work |
| ADH20 | ΔvirB4::Km (polar) | This work |
| ADH21 | ΔvirB4 (nonpolar) | This work |
| ADH22 | ΔvirB5::Km (polar) | This work |
| ADH23 | ΔvirB5 (nonpolar) | This work |
| ADH24 | ΔvirB6::Km (polar) | This work |
| ADH25 | ΔvirB6 (nonpolar) | This work |
| ADH26 | ΔvirB7::Km (polar) | This work |
| ADH27 | ΔvirB7 (nonpolar) | This work |
| ADH28 | ΔvirB8::Km (polar) | This work |
| ADH29 | ΔvirB8 (nonpolar) | This work |
| ADH30 | ΔvirB9::Km (polar) | This work |
| ADH31 | ΔvirB9 (nonpolar) | This work |
| ADH32 | ΔvirB10::Km (polar) | This work |
| ADH33 | ΔvirB10 (nonpolar) | This work |
| ADH15 | ΔvirB11::Km (polar) | This work |
| ADH16 | ΔvirB11 (nonpolar) | This work |
| AK/ORF12 | ΔvirB12::Km | 59 |
| E. coli | ||
| DH5α | endA1 hsdR17(rK− mK−) supE44 thi-1 recA1 gyrA relA1 Δ(lacZYA-argF)U169 deoR φ80dlacZΔ(M15) | 65 |
| BL21(DE3) | F−ompT gal [dcm] [lon] hsdSB(rB− mB−) DE3 | 58 |
| Plasmids | ||
| pBluescript KS | colE1, bla | Stratagene |
| pCR2.1-TOPO | TA cloning vector | Invitrogen |
| pUC4-KSAC | Carries Tn903 kanamycin resistance cassette | Pharmacia |
| pUC4-KIXX | Carries Tn5 kanamycin resistance cassette | Pharmacia |
| pEX18Ap | sacB | 28 |
| pBBRmcs4 | ampR | 33 |
| pADH100 | virB 940-1939 and virB 2290-3290 cloned into pCR2.1-TOPO | This work |
| pADH101 | virB 940-1939 and virB 2290-3290 separated by a kanamycin resistance cassette KIXX cloned into pCR2.1-TOPO | This work |
| pADH102 | SacB cloned into pADH100 | This work |
| pADH103 | virB 1290-2289 and virB 4786-5785 cloned into pCR2.1-TOPO | This work |
| pADH104 | virB 1290-2289 and virB 4786-5785 separated by kanamycin resistance cassette KIXX cloned into pCR2.1-TOPO | This work |
| pADH105 | sacB cloned into pADH103 | This work |
| pADH106 | virB 2772-4707 and virB 5507-6506 cloned into pBluescript | This work |
| pADH107 | virB 2772-4707 and virB 5507-6506 separated by kanamycin resistance cassette KSAC cloned into pBluescript | This work |
| pADH108 | sacB cloned into pADH106 | This work |
| pADH109 | virB 4690-5689 and virB 6734-7751 cloned into pBluescript | This work |
| pADH110 | virB 4690-5689 and virB 6734-7751 separated by kanamycin resistance cassette KSAC cloned into pBluescript | This work |
| pADH111 | sacB cloned into pADH109 | This work |
| pADH112 | virB 5897-6896 and virB 7071-8070 cloned into pBluescript | This work |
| pADH113 | virB 5897-6896 and virB 7071-8070 separated by a kanamycin resistance cassette, KSAC, cloned into pBluescript | This work |
| pADH114 | sacB cloned into pADH112 | This work |
| pADH115 | virB 6073-7072 and virB 7786-8637 cloned into pCR2.1-TOPO | This work |
| pADH116 | virB 6073-7072 and virB 7786-8637 separated by a kanamycin resistance cassette, KIXX, cloned into pCR2.1-TOPO | This work |
| pADH117 | sacB cloned into pADH115 | This work |
| pADH118 | virB 6789-7788 and virB 8655-9658 cloned into pBluescript | This work |
| pADH119 | virB 6789-7788 and virB 8655-9658 separated by a kanamycin resistance cassette, KSAC, cloned into pBluescript | This work |
| pADH120 | sacB cloned into pADH118 | This work |
| pADH121 | virB 7654-8654 and virB 9802-10819 cloned into pBluescript | This work |
| pADH122 | virB 7654-8654 and virB 9802-10819 separated by a kanamycin resistance cassette, KSAC, cloned into pBluescript | This work |
| pADH123 | sacB cloned into pADH121 | This work |
| pADH124 | virB 8351-9822 and virB 10850-12441 cloned into pBluescript | This work |
| pADH125 | virB 8351-9822 and virB 10850-12441 separated by kanamycin resistance cassette KSAC cloned into pBluescript | This work |
| pADH126 | SacB cloned into pADH124 | This work |
| pVP | 450-bp fragment of virB promoter cloned into pBBR-MCS4 | This work |
| pVP3 | virB3 cloned into pVP | This work |
| pVP4 | virB4 cloned into pVP | This work |
| pVP5 | virB5 cloned into pVP | This work |
| pVP6 | virB6 cloned into pVP | This work |
| pVP7 | virB7 cloned into pVP | This work |
| pVP8 | virB8 cloned into pVP | This work |
| pVP9 | virB9 cloned into pVP | This work |
| pVP10 | virB10 cloned into pVP | This work |
| pVP11 | virB11 cloned into pVP | This work |
a
Cloned fragments of the virB operon are numbered according to the sequence found under GenBank accession number AF226278 (56).