TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYZ-argF) recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 deoR U169 | Gibco BRL |
| SM10 | Mobilizing strain, RP4 tra genes integrated in chromosome, Kmr | 52 |
| XL1-Blue MR | Nonmethylating (dam dcm) cloning host used for cosmid library construction and preparation of DNA for electroporation into B. cepacia: Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac |
Stratagene |
| B. cepacia | ||
| K56-2 | Clinical isolate, genomovar III | 11, 27 |
| K56-I117 | pvdA::Tn5-OT182 derivative of K56-2 | This study |
| K56-T10 | pvdD::Tn5-OT182 derivative of K56-2 | This study |
| K56pvdA::tp | pvdA::Tpr derivative of K56-2 | This study |
| Plasmids | ||
| pOT182 | pSUP102(GM)::Tn5-OT182; Cmr Gmr Apr Tcr | 33 |
| pNOT19 | Modified pUC19 cloning vector, Apr | 47 |
| pUCP28T | Broad-host-range vector; IncP OriT; pRO1600 ori; Tpr | 50 |
| pPDI117-2 | 9.5-kb EcoRI fragment from I117 obtained by self-cloning, Tcr | This study |
| pPDI117-3 | 11.5-kb NotI fragment from I117 obtained by self-cloning, Tcr | This study |
| pPD519 | pNOT19 with 6.1-kb SphI fragment from K56-2 with pvdA gene | This study |
| pPD520 | pUCP28T with 6.5-kb SphI fragment from pPD519 | This study |
| p34E-Tp | Source of Tp cassette; Tpr | 13 |
| pRK2013 | ColE1 Tra (RK2)+ Kmr | 16 |
| pEX18Tc | Suicide vector; sacB Tcr | 23 |
| pZ1918 | Apr Gmr; source of promoterless lacZ gentamicin resistance cassette | 48 |
| pPD520-6Z | pPD520 with 4.3-kb lacZ-Gmr cassette from pZ1918 in BglII site of pvdA gene; same orientation | This study |
| pPD520-14Z | pPD520 with 4.3-kb lacZ-Gmr cassette from pZ1918 in BglII site of pvdA gene; reverse orientation | This study |
| pUC-TP | pUC-GM (46) with the gentamicin resistance cassette replaced with a 1.1-kb Tpr gene cassette; Apr Tpr |
This study |
| pScosBC1 | Novel broad-host-range cosmid cloning vector based on pSuperCos1 (67); Apr Tpr | This study |
| pSBC-4 | pScosBC1 from K56-2 library containing the pvdA gene | This study |
| pSBC-8 | pScosBC1 from K56-2 library containing the pvdA gene | This study |
| pSBC-13 | pScosBC1 from K56-2 library containing the pvdA gene | This study |