TABLE 1.
Bacterial strains and plasmids developed in this study
| Xanthomonas strain or plasmid | Descriptiona |
|---|---|
| X. axonopodis pv. vesicatoria | |
| 7882.3 | Cus, Rifr, Kanr; kanamycin cassette was inserted into the SalI site of copL in 7882 (10) |
| 78518.2 | Cur Rifr; the Cur plasmid of 7882 was electroporated into strain 78518 (10) |
| Plasmids | |
| pCOP151 | Smr Cmr; 1.9-kb BamHI fragment of pCOP120 (55) cloned in the BglII site of pMP190 (49) in the correct orientation to express lacZ |
| pCOP153B | Apr; 1.9-kb BamHI fragment of pCOP116 (55) cloned in pUC128 (25) in the anti-lac orientation |
| pCOP156 | Apr; pCOP153B was digested with KpnI to result in a deletion of the 500-bp KpnI-BamHI fragment and then relegated |
| pCOP157 | Apr; pCOP153B was digested with ClaI to result in a deletion of the 1.4-kb ClaI-BamHI fragment and then relegated |
| pCOP161 | Smr Cmr; 1.5-kb XbaI-KpnI-fragment of pCOP156 cloned in pMP190 |
| pCOP162 | Smr Cmr; 510-bp XbaI-KpnI fragment of pCOP157 cloned in pMP190 |
| pCOP166 | Apr; pCOP153B was digested with SphI to result in a deletion of 0.6-kb SphI-BamHI fragment and then religated |
| pCOP168 | Apr; pCOP156 was digested with SacII to result in a deletion of the 440-bp BamHI-SacII fragment and then religated |
| pCOP169 | Smr Cmr; 1.0-kb (SacII)b-Xbal fragment of pCOP168 cloned in (SalI)-XbaI sites of pMP190 |
| pCOP170 | Smr Cmr; 1.0-kb (XbaI)-SphI fragment of pCOP166 cloned in (XbaI)-KpnI sites of pMP190 |
| pCOP187 | Tcr; 510-bp BamHI-HindIII fragment of pCOP157 cloned in pRK415 (25) |
| pCOP189 | Tcr; 8.8-kb HindIII fragment of pDAC102 (34) cloned in the HindIII site of pCOP187, resulting in transcriptional fusion of the constitutive promoter with copABCDRS. |
| pCOP191 | Apr; pCOP153B was digested with ApaI to result in a deletion of 900-bp ApaI-BamHI fragment and then religated |
| pCOP192 | Apr; pCOP191 digested with BamHI and StuI, blunt ended, and religated |
| pCOP193 | Apr; pCOP191 digested with SalI and ClaI, blunt ended, and religated |
| pCOP196 | Apr; pCOP191 digested with StuI and ApaI, blunt ended and relegated |
| pCOP197 | Smr Cmr; the 985-bp XbaI-KpnI fragment of PCOP191 cloned in pMP190 |
| pCOP198 | Smr Cmr; the 735-bp XbaI-KpnI fragment of pCOP192 cloned in pMP190 |
| pCOP199 | Smr Cmr; the 935-bp Xbal-KpnI fragment of pCOP193 cloned in pMP190 |
| pCOP202 | Smr; Cmr; the 250-bp Xbal-KpnI fragment of pCOP196 cloned in pMP190 |
| pCOP203 | Apr; the 470-bp ClaI-ApaI fragment of pCOP191 cloned in pUC128 |
| pCOP204 | Apr; the 200 bp StuI-SalI fragment of pCOP191 cloned into the SmaI-SalI sites of pUC128 |
| pCOP205 | Smr Cmr; the 470-bp XbaI-KpnI fragment of pCOP203 cloned in pMP190 |
| pCOP206 | Smr Cmr; the 200-bp XbaI-KpnI fragment of pCOP204 cloned in pMP190 |
| pCOP210 | Apr; pCOP192 was digested with ClaI blunt ended with Klenow and religated |
| pCOP214 | Smr; Cmr; the 725-bp XbaI-KpnI fragment of pCOP210 was cloned in pMP190 |
| pCOPM1 | Smr; Cmr; substitution of first Met codon in copL (in pCOP192) to Val and subcloned into pMP190 |
| pCOPM2 | Smr; Cmr; substitution of second Met codon in copL (in pCOP192) to Leu and subcloned into pMP190 |
| pCOPM3 | Smr; Cmr; substitution of third Met codon in copL (in pCOP192) to Leu and subcloned into pMP190 |
a
Cur, copper resistance; Cus, copper sensitivity; Rifr, rifampin resistance; Kmr, kanamycin resistance; Smr, streptomycin-resistance; Apr, ampicillin resistance; Tcr, tetracycline resistance; Cmr, chloramphenicol resistance.
b
Restriction sites in parentheses describe sites that have been blunt ended before ligation.