TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 Δ(lac)U169 (φ80dlacΔM15) | 3 |
| XL1-Blue MR | Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac | Stratagene |
| S17-1 | thi pro recA hsdR Tpr Smr; chromosomally integrated RP4-Tc::Mu-Km::Tn7 | 34 |
| JM109(λpir) | λpir lysogen of JM109 | Lab collection |
| B. multivorans | ||
| ATCC 17616 | Soil isolate; genomovar II of the Bcc | ATCCb |
| BT141 | ATCC 17616::TnMod-RTp′; leaky Trp− | This study |
| BT211 | ATCC 17616 leuD::kana; knockout mutant by use of pKOM154 | This study |
| BT214 | ATCC 17616 Δ(trpF-B)::kan; knockout mutant by use of pKOM157 | This study |
| BT216 | ATCC 17616 with the insert of the TnMod-RTp′-Ωkan cassette from pKOM160 | This study |
| Plasmids | ||
| pTnMod-RTp′ | Tpr; TnMod-RTp′-containing plasmid whose transposable region containing the Tpr gene, R6KoriV, and PacI and SwaI sites is able to transpose by the transposase that is encoded outside of the TnMod-RTp′ region | 12 |
| pUC18 | Apr; cloning vector | 41 |
| pUC19 | Apr; cloning vector | 41 |
| pUC4K | KmrApr; kan flanked by inverted repeats of oligonucleotide containing SalI and PstI sites | 36 |
| pEX18Tc | TcrsacB oriT; suicide vector for gene replacement carrying the pUC18-derived multiple cloning sites | 18 |
| pHP45ΩKm | Apr Kmr; Ωkan flanked by transcription terminators and BamHI sites | 13 |
| pBBR1MCS | Cmr; broad-host-range cloning vector | 20 |
| SuperCosl | Apr Kmr; cosmid vector | Stratagene |
| pSBm1-92 | SuperCos1 derivative carrying the trpB region of ATCC 17616 | This study |
| pBTB141 | Tpr; TnMod-RTp′-containing plasmid recovered by self-ligation of the BamHI-treated genome of BT141 | This study |
| pKOM152 | pEX18Tc derivative carrying the BamHI-linearized form of pBTB141 | This study |
| pKOM154 | pEX18Tc derivative with the pKOM152-derived, leuD-containing BamHI-EcoRI fragment that has the insert of the pUC4K-derived kan in leuD | This study |
| pKOM157 | pEX18Tc derivative with the trpFB-containing BamHI fragment in which the XhoI fragment containing these genes is replaced by the SalI-flanked kan from pUC4K | This study |
| pKOM159 | pEX18Tc derivative with the TnMod-RTp′-containing EcoRI fragment from pKOM152 | This study |
| pKOM160 | pKOM159 derivative in which the TnMod-RTp′-derived BglII site has the insert of the BamHI-flanked Ωkan from pHP45ΩKm | This study |
a
kan, Kmr gene.
b
ATCC, American Type Culture Collection.