Table 1.
Strains, plasmids, and oligonucleotides used in this study
| Strain, plasmid, or oligonucleotide | Description or sequence (5′–3′)a | Source or reference |
|---|---|---|
| Strains | ||
| A. mimigardefordensis DPN7 | Wild type, DTDP-degrading bacterium | 1 (DSM 17166T, LMG 22922T) |
| A. mimigardefordensis ΔsucCD | Deletion of sucCD, no growth on DTDP and 3SP | This study |
| E. coli Top10 | F−mcrAΔ(mrr-hsdRMS-mcrBC) rpsL nupG φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK endA1 | Invitrogen, Carlsbad, CA |
| E. coli BL21(DE3)/pLysS | F−ompT hsdSB (rB− mB−) gal dcm (DE3), pLysS (Cmr) | Novagen, Madison, WI |
| E. coli BL21(DE3) | F−ompT hsdSB (rB− mB−) gal dcm (DE3) | Novagen, Madison, WI |
| Plasmids | ||
| pSUP5011 | Apr Cmr Kmr Tn5::mob | 27 |
| pBluescript SK(–) | AprlacPOZ′ | Stratagene, San Diego, CA |
| pBluescriptSK::sucCDrbs | Apr | This study |
| pGEM-T Easy | AprlacPOZ′ | Promega, Madison, WI |
| pGEM-T Easy::sucCDrbs | AprlacPOZ′ | This study |
| pJQ200mp18Tc | TcrsacB oriV oriT traJ | 35 |
| pJQ200mp18Tc::ΔsucCD | TcrsacB oriV oriT traJ | This study |
| Oligonucleotides | ||
| M13 forward | GTAAAACGACGGCCAGT | MWG Biotech AG, Ebersberg, Germany |
| M13 reverse | CAGGAAACAGCTATGAC | MWG Biotech AG, Ebersberg, Germany |
| IS50 walking | TCGGCCGCACGATGAAGAGC | MWG Biotech AG, Ebersberg, Germany |
| IS50 sequencing | CGTTACCATGTTAGGAGGTCACATGG | MWG Biotech AG, Ebersberg, Germany |
| sucCDforward_PstI | CTGCAGCAGTCTCAATTCGTGTGCTCGC | MWG Biotech AG, Ebersberg, Germany |
| sucCDreverse_XhoI_stop | CTCGAGTTACAGTACTGATTTGAGCAGTTTG | MWG Biotech AG, Ebersberg, Germany |
| sucCXbaI | TCTAGATGTCTCTGGGTTCTTCGGCAC | MWG Biotech AG, Ebersberg, Germany |
| sucCEcoRI | GAATTCGTATTACCTATTAACGTAGGATAAAAAAAC | MWG Biotech AG, Ebersberg, Germany |
| sucDXbaI | TCTAGACGCCTTCATCGTCGTCTGAC | MWG Biotech AG, Ebersberg, Germany |
| sucDEcoRI | AAAAGAATTCGTACCAGCGGTCATTTGTCG | MWG Biotech AG, Ebersberg, Germany |
a
For the abbreviations used in the E. coli genotypes, see reference 3. Cmr, chloramphenicol resistance; Apr, ampicillin resistance; Kmr, kanamycin resistance; Tcr, tetracycline resistance.