TABLE 1.
Strains and plasmids
| Strain or plasmid | Characteristic(s)a | Source or reference |
|---|---|---|
| Strains | ||
| E. coli HB101 | F− Δ(mcrC-mrr) recA13 rpsL20 | Laboratory stock |
| B. fragilis TM4000 | Clinical isolate, Rifr, aerotolerant | 8 |
| B. fragilis Tn4400′ mutants | ||
| JD3a.2 | Arginine auxotroph | This study |
| JD3a.3 | Arginine auxotroph | This study |
| JD4.9 | Arginine auxotroph | This study |
| YT44.2.1 | Tryptophan auxotroph | This study |
| YT58.1.3 | MGD bafA mutant | This study |
| YT65.2.10 | MGD, less aerotolerant than TM4000 | This study |
| YT112.2.16 | MGD | This study |
| YT118.1.13 | MGD | This study |
| YT120.1.7 | MGD | This study |
| YT123.2.18 | MGD | This study |
| YT129.2.20 | MGD, less aerotolerant than TM4000 | This study |
| YT135.2.8 | MGD, less aerotolerant than TM4000 | 13 |
| Plasmids | ||
| pDG5 | Ampr, E. coli plasmid carrying oriT from RP4 | 4 |
| pFD544 | E. coli replicon, tetQ gene | 6 |
| pJST51 | pDG5 with endR1 gene from E. coli | 14 |
| pNJR609 | BglII fragment containing Tn4400 from pOX446R1 cloned into pDG5 | This study |
| pNJR609Δ | pNJR609 with a spontaneous deletion, Ampr Tetr in E. coli | This study |
| pOX446R1 | Derived from pOX446 by homologous recombination, contains a single copy of Tn4400 | 7 |
| pRG23 | SacI tetQ fragment from pFD544 cloned into pSP72 | This study |
| RK231 | RP4 derivative, Kanr Tetr Tra+ cloning vector, Ampr | 4 |
| pSP72 | Promega, Madison, Wis. | |
| pYT644A and -B | oriT of RP4 from pJST51 cloned into the PstI site of pNJR609Δ, Amps Tetr (pYT644A and -B differ in the orientation of oriT) | This study |
| pYT645A and -B | tetQ is cloned as a BglII-BamHI fragment into the unique BglII sites of pYT644A and pYT644B | This study |
| pYT646A and -B | 1.6-kb PstI fragment from pJST51 cloned into the PstI sites of pYT645A and pYT645B, restoring the bla gene; Ampr Tetr in E. coli | This study |
a
Rifr, rifampin resistant; Kanr, kanamycin resistant; Tra+, transfer proficient.