TABLE 2.
Plasmids used in this study
| Plasmid | Description | Purpose | Antibiotic resistance | Reference or source |
|---|---|---|---|---|
| pBluescript II KS | High-copy-number cloning vector | Initial subcloning | Apr | Stratagene |
| pBluescript II KS-Km | HindIII/SmaI-digested wild-type Tn5 treated with the Klenow fragment and ligated to EcoRV-digested pBluescript II KS | Source of aphA gene | Apr Kmr | This study |
| pGP704 | oriR6K mobRP4 | Suicide vector | Apr | 37 |
| pGP704Km | aphA gene inserted into the bla gene of pGP704 | Suicide vector | Kmr | This study |
| pACYC184 | Low-copy-number cloning vector | Complementation | Tcr Cmr | 7 |
| pUC18CML | High-copy-number cloning vector | Source of cat gene | Apr Cmr | 51 |
| pBR322 | Low-copy-number cloning vector | Complementation | Tcr | |
| pRAB2401 | Subclone of O75 rfb gene cluster | Subcloning of rfb genes | Apr | 4 |
| pSMB102 | Intramolecular ligation of pRAB2401 cut with PstI | Cloning of rfbD gene | Apr | This study |
| pSMB103 | cat gene (SmaI/HindII) inserted into SnaBI of pSMB102 | Mutation of rfbD gene | Apr Cmr | This study |
| pSMB104 | EcoRV fragment of pSMB103 ligated to pGP704Km | Subcloning into suicide vector for conjugation | Kmr Cmr | This study |
| pSMB105 | Intramolecular ligation of pRAB2401 cut with ClaI | Cloning of rfbKM genes | Apr | This study |
| pSMB106 | 6.5-kb ClaI/KpnI fragment of pSMB105 inserted into pBluescript II KS | Subcloning of rfbKM genes | Apr | This study |
| pSMB107 | Deletion of 1.1-kb NruI fragment from rfbKM and insertion of cat gene (SmaI/HindII) | Mutation of rfbKM genes | Apr Cmr | This study |
| pSMB108 | EcoRV/KpnI fragment of pSMB107 ligated to pGP704 | Subcloning into suicide vector for conjugation | Apr Cmr | This study |
| pSMB109 | MunI fragment of pSMB106 ligated to pBR322 cut with EcoRI/ScaI | Cloning of rfbKM genes for complementation | Tcr | This study |
| pSMB110 | ugd PCR product (EcoRV/BamHI) ligated to pBluescript II KS | Cloning of sequence upstream of rol gene | Apr | This study |
| pSMB111 | his PCR product (BamHI/XbaI) ligated to pSMB110 | Cloning of upstream and downstream sequences of rol gene | Apr | This study |
| pSMB112 | Insertion of cat gene (BamHI ends) into BamHI site of pSMB111 | cat gene inserted between ugd and his | Apr Cmr | This study |
| pSMB114 | EcoRV/SalI fragment of pSMB112 ligated to pGP704 | Subcloning into suicide vector for conjugation | Apr Cmr | This study |
| pGH1610 | galF rfbBDA′04 from pGH161 (20) cut with PstI/HpaI and ligated to pBR322 cut with PstI/ScaI | Cloning of rfbD gene for complementation | Tcr | This study |
| pRAB10 | rol gene in pACYC184 | Subcloning of rol gene for complementation | Tcr | 3 |
| pSMB250 | kfiCD PCR product (SalI ends) ligated to pBluescript II KS | Cloning of kfiCD genes | Apr | This study |
| pSMB251-Cm | Insertion of cat gene (SmaI/HindII) into NdeI-filled ends of pSMB250 | Mutation of kfiC gene | Apr Cmr | This study |
| pSMB251-Km | Insertion of aphA gene (filled ends) into NdeI-filled ends of pSMB250 | Mutation of kfiC gene | Apr Kmr | This study |
| pSMB252-Cm | SalI fragment of pSMB251-Cm ligated to pGP704 | Subcloning into suicide vector for conjugation | Apr Cmr | This study |
| pSMB252-Km | SalI fragment of pSMB251-Km ligated to pGP704 | Subcloning into suicide vector for conjugation | Apr Kmr | This study |