Table 1.

Strains and plasmids used in this studya

Strain or plasmidRelevant characteristicsReference or source
E. coliDH5αMCRRecA GnS 6
B. thetaiotaomicron
 5482Wild type, Gnr Anaerobe Laboratory, Virginia Polytechnic Institute, Blacksburg
 4007Wild type, Tetr GnrG7+ Am+ B. thetaiotaomicron 5482 (Rifr) with CTn DOT
 Ms-4EmrGnr G7+Am Tn4351-generated mutant of strain 5482 (2)
 ΩsusA EmrGnr G7+Am+ Bacteroides suicide vector pGERM containing a PCR-generated 0.65-kbp KpnI-BamHI fragment inserted in B. thetaiotaomicron chromosome insusA (4)
 ΩsusB Tetr GnrG7 Am Bacteroides suicide vector pBT-1 containing a PCR-generated 0.61-kbpKpnI-BamHI fragment inserted in the B. thetaiotaomicron chromosome in susB (pBT1-SB) (4)
 ΩsusC TetrGnr G7Am Bacteroides suicide vector pBT-1 containing a PCR-generated 0.61-kbp KpnI-BamHI fragment inserted in the B. thetaiotaomicron chromosome insusC (pBT1-SC) (15)
 ΩsusG Tetr GnrG7+ Am Bacteroides suicide vector pBT-1 containing a PCR-generated 0.61-kbpKpnI-BamHI fragment inserted in the B. thetaiotaomicron chromosome in susG (pBT1-SC) (15)
 ΩSABEmr TetrGnr G7 Am Double-insertion strain derived from ΩsusA in which pBT1-SB was used to make an insertion in susB (this study)
 ΩSACEmr Tetr GnrG7 Am Double-insertion strain derived from ΩsusA in which pBT1-SC was used to make an insertion in susC (this study)
 ΩSAGEmrTetr Gnr G7+Am Double-insertion strain derived from ΩsusA in which pBT1-SG was used to make an insertion insusG (this study)
Plasmids
 pBT-1Knr (Tcr)RSF1010-based suicide vector used to make insertional disruptions (19)
 pGERMApr (Ermr)pUC19-based suicide vector expressing ermG used to make insertions in the B. thetaiotaomicron chromosome (17; this study)
 pNLY1::PsusAAprCmr (Cmr)pACYC-based shuttle vector containing PsusA used to express genes in trans(this study)
 pSGC23AApr Cmr(Cmr)pNLY1::PsusA containing a PCR-generatedsusG 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 inE. coli; antibiotic resistances in parentheses are expressed only in B. thetaiotaomicron.