TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Characteristicsa | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 deoR recA1 hsdR17(rK− mK+) phoA supE44 λ−thi-1 gyrA96 relA1 | Gibco-BRL |
| BL21(DE3) | F−ompT hsdSB(rB− mB−) gal dcm (DE3) | Novagen |
| Xanthomonas campestris pv. vesicatoria | ||
| 82-8 | Wild type, race 1 | R. E. Stall |
| E-3 | Wild type, race 2 | R. E. Stall |
| LS833 | Wild type, race 3 | S. Lee |
| Plasmids | ||
| pT7-7 | T7 promoter-based expression vector; Ampr | 24 |
| pET14b | T7 promoter-based expression vector; Ampr | Novagen |
| pLysS | Encoding T7 lysozyme gene; Cmr | Novagen |
| pBluescript II SK(+) | Phagemid, pUC derivative; Ampr | Stratagene |
| pBSN | pBluescript II SK(+) with NdeI site in β-galactosidase gene start codon | This study |
| pBHPAG | 0.4-kb NdeI-BamHI fragment, including entire hpaG gene, cloned into pBSN | This study |
| pHpaGMT | SmaI-BssHII fragment (T7 promoter region) of pBHPAG was deleted and religated; template for site-directed mutagenesis of hpaG | This study |
| pTJ1 | PCR-amplified hpaG in pT7-7; Ampr | This study |
| pJ14 | hpaG in pET14b; Ampr | This study |
| pHGM61 | hpaG(L4A) in pET14b; Ampr | This study |
| pHGM62 | hpaG(Q7A) in pET14b; Ampr | This study |
| pHGM63 | hpaG(G9A) in pET14b; Ampr | This study |
| pHGM64 | hpaG(F14D) in pET14b; Ampr | This study |
| pHGM65 | hpaG(Q16A) in pET14b; Ampr | This study |
| pHGM66 | hpaG(P19A) in pET14b; Ampr | This study |
| pHGM67 | hpaG(G20E) in pET14b; Ampr | This study |
| pHGM68 | hpaG(Q21A) in pET14b; Ampr | This study |
| pHGM69 | hpaG(Q24A) in pET14b; Ampr | This study |
| pHGM70 | hpaG(Q29A) in pET14b; Ampr | This study |
| pHGM71 | hpaG(G30E) in pET14b; Ampr | This study |
| pHGM72 | hpaG(Q32A) in pET14b; Ampr | This study |
| pHGM73 | hpaG(G33A) in pET14b; Ampr | This study |
| pHGM26 | hpaG(S35A) in pET14b; Ampr | This study |
| pHGM7 | hpaG(E36A) in pET14b; Ampr | This study |
| pHGM27 | hpaG(K37A) in pET14b; Ampr | This study |
| pHGM28 | hpaG(Q38A) in pET14b; Ampr | This study |
| pHGM46 | hpaG(L39A) in pET14b; Ampr | This study |
| pHGM74 | hpaG(L39P) in pET14b; Ampr | This study |
| pHGM6 | hpaG(D40A) in pET14b; Ampr | This study |
| pHGM29 | hpaG(Q41A) in pET14b; Ampr | This study |
| pHGM46 | hpaG(L42A) in pET14b; Ampr | This study |
| pHGM75 | hpaG(L42D) in pET14b; Ampr | This study |
| pHGM36 | hpaG(L43P) in pET14b; Ampr | This study |
| pHGM30 | hpaG(T44A) in pET14b; Ampr | This study |
| pHGM40 | hpaG(Q45A) in pET14b; Ampr | This study |
| pHGM32 | hpaG(L46A) in pET14b; Ampr | This study |
| pHGM41 | hpaG(I47A) in pET14b; Ampr | This study |
| pHGM11 | hpaG(L50A) in pET14b; Ampr | This study |
| pHGM76 | hpaG(L50P) in pET14b; Ampr | This study |
| pHGM12 | hpaG(L51A) in pET14b; Ampr | This study |
| pHGM13 | hpaG(Q52A) in pET14b; Ampr | This study |
| pHGM42 | hpaG(Q53A) in pET14b; Ampr | This study |
| pHGM15 | hpaG(S54A) in pET14b; Ampr | This study |
| pHGM16 | hpaG(N55A) in pET14b; Ampr | This study |
| pHGM18 | hpaG(E58A) in pET14b; Ampr | This study |
| pHGM8 | hpaG(Q59A) in pET14b; Ampr | This study |
| pHGM19 | hpaG(G60V) in pET14b; Ampr | This study |
| pHGM9 | hpaG(Q61A) in pET14b; Ampr | This study |
| pHGM20 | hpaG(G62V) in pET14b; Ampr | This study |
| pHGM21 | hpaG(Q63A) in pET14b; Ampr | This study |
| pHGM38 | hpaG(G64A) in pET14b; Ampr | This study |
| pHGM10 | hpaG(Q65A) in pET14b; Ampr | This study |
| pHGM22 | hpaG(G66A) in pET14b; Ampr | This study |
| pHGM23 | hpaG(G67V) in pET14b; Ampr | This study |
| pHGM5 | hpaG(D99A) in pET14b; Ampr | This study |
| pHGM17 | hpaG(I120T) in pET14b; Ampr | This study |
| pHGM37 | hpaG(L121P) in pET14b; Ampr | This study |
| pHGM2 | hpaG(A126T) in pET14b; Ampr | This study |
| pHGM34 | hpaG(A126V) in pET14b; Ampr | This study |
| pHGM3 | hpaG(S131L) in pET14b; Ampr | This study |
| pHGM77 | hpaG(L43P/L50P) in pET14b; Ampr | This study |
| pTHGC | hpaG(ΔN67) in pT7-7; Ampr | This study |
| pTHG67 | hpaG(ΔC66) in pET14b; Ampr | This study |
| pTHG58 | hpaG(ΔC75) in pET14b; Ampr | This study |
| pTHG56 | hpaG(ΔC77) in pET14b; Ampr | This study |
| pTHG54 | hpaG(ΔC79) in pET14b; Ampr | This study |
| pTHG52 | hpaG(ΔC81) in pET14b; Ampr | This study |
| pTHG50 | hpaG(ΔC83) in pET14b; Ampr | This study |
| pTXOPA | PCR product of xopA cloned into pET14b; Ampr | This study |
| pTXH2 | 120-bp NdeI-PvuII fragment of pTXOPA and 73-bp PvuII-BamHI fragment of pTHG67 coligated into pET14b; Ampr | This study |
| pTXM1 | xopA(F48L) in pET14b; Ampr | This study |
| pTXM2 | xopA(M52L) in pET14b; Ampr | This study |
| pTXM4 | xopA(F48L/M52L) in pET14b; Ampr | This study |
a
Ampr, ampicillin resistance; Cmr, chloramphenicol resistance.