TABLE 1.
Strains and plasmids used in this studya
| Strain or plasmid | Relevant characteristics | Reference or source |
|---|---|---|
| E. coli DH5αMCR | RecA GnS | 6 |
| B. thetaiotaomicron | ||
| 5482 | Wild type, Gnr | Anaerobe Laboratory, Virginia Polytechnic Institute, Blacksburg |
| 4007 | Wild type, Tetr Gnr G7+ Am+ | B. thetaiotaomicron 5482 (Rifr) with CTn DOT |
| Ms-4 | Emr Gnr G7+ Am− | Tn4351-generated mutant of strain 5482 (2) |
| ΩsusA | Emr Gnr G7+ Am+ | Bacteroides suicide vector pGERM containing a PCR-generated 0.65-kbp KpnI-BamHI fragment inserted in B. thetaiotaomicron chromosome in susA (4) |
| ΩsusB | Tetr Gnr G7− Am− | Bacteroides suicide vector pBT-1 containing a PCR-generated 0.61-kbp KpnI-BamHI fragment inserted in the B. thetaiotaomicron chromosome in susB (pBT1-SB) (4) |
| ΩsusC | Tetr Gnr G7− Am− | Bacteroides suicide vector pBT-1 containing a PCR-generated 0.61-kbp KpnI-BamHI fragment inserted in the B. thetaiotaomicron chromosome in susC (pBT1-SC) (15) |
| ΩsusG | Tetr Gnr G7+ Am− | Bacteroides suicide vector pBT-1 containing a PCR-generated 0.61-kbp KpnI-BamHI fragment inserted in the B. thetaiotaomicron chromosome in susG (pBT1-SC) (15) |
| ΩSAB | Emr Tetr Gnr G7− Am− | Double-insertion strain derived from ΩsusA in which pBT1-SB was used to make an insertion in susB (this study) |
| ΩSAC | Emr Tetr Gnr G7− Am− | Double-insertion strain derived from ΩsusA in which pBT1-SC was used to make an insertion in susC (this study) |
| ΩSAG | Emr Tetr Gnr G7+ Am− | Double-insertion strain derived from ΩsusA in which pBT1-SG was used to make an insertion in susG (this study) |
| Plasmids | ||
| pBT-1 | Knr (Tcr) | RSF1010-based suicide vector used to make insertional disruptions (19) |
| pGERM | Apr (Ermr) | pUC19-based suicide vector expressing ermG used to make insertions in the B. thetaiotaomicron chromosome (17; this study) |
| pNLY1::PsusA | Apr Cmr (Cmr) | pACYC-based shuttle vector containing PsusA used to express genes in trans (this study) |
| pSGC23A | Apr Cmr (Cmr) | pNLY1::PsusA containing a PCR-generated susG gene cloned downstream of PsusA (this study) |
a
Abbreviations: G7, maltoheptaose; Am, amylopectin; Ap, ampicillin; Cm, chloramphenicol; Em, erythromycin; Gn, gentamicin; Tc, tetracycline; CTn, conjugative transposon; CTn DOT, conjugative transposon which contains the tetQ gene. Antibiotic resistances not in parentheses are expressed only in E. coli; antibiotic resistances in parentheses are expressed only in B. thetaiotaomicron.