TABLE 1.
Bacterial strains, plasmids, and oligonucleotides used in this work
| Strain, plasmid, or oligonucleotide | Relevant characteristics | Source or reference |
|---|---|---|
| R. capsulatus strains | ||
| MT1131 | Parental strain | F. Daldal |
| DL1 | thyX::Specr | This study |
| R. sphaeroides Ga | crt | F. Daldal |
| E. coli strains | ||
| SURE | e14− (McrA−) Δ(mcrCB-hsdSMR-mrr)171 endA1 supE44 thi-1 gyrA96 relA1 lac recB recJ sbcC umuC::Tn5 (Kanr) uvrC [F′ proAB lacIqZΔM15 Tn10 (Tetr)] | Stratagene |
| HB101 | F−proA2 hsdS20 recA13 ara-14 lacY xyl-5 galK2 rpsL20 supE44 rplsL20 supE44 proA2 mtl-1 | ATCC |
| S17-1 | thi pro ara recA StrepRP4 2-Tet::Mu-Kan::Tn7 | 27 |
| Plasmids | ||
| pGEM-T Easy | Ampr | Promega |
| pBluescript II KS+ | Ampr | Stratagene |
| pRK415 | Tetr broad-host-range plasmid | 13 |
| pRK2013 | Kanr helper | 6 |
| pHP45Ω | Specr Ampr | 22 |
| pSUP202 | Ampr Cmr Tetr, suicide plasmid | 27 |
| pDL5 | R. capsulatus thyX in pGEM-T Easy | This study |
| pDL7 | 3.5-kb PstI fragment with thyX::Specr allele in pSUP202 | This study |
| pDL9 | 1.6-kb NotI-BglII fragment with thyX gene in pBluescript II KS+ | This study |
| pDL11 | 1.4-kb PstI fragment with R. capsulatus thyX gene in pRK415 | This study |
| pDL12 | 1.7-kb SpeI-EcoRI fragment with R. sphaeroides thyA and folA in pRK415 | This study |
| pDL14 | 1.4-kb XbaI-EcoRI fragment with R. sphaeroides thyA in pRK415 | This study |
| Oligonucleotides | ||
| ORF311-A | 5′-ATGTCGCTTACCCCAGACC | This study |
| ORF311-B | 5′-ACCCCACTCCTCCACAAACT | This study |
| ORF311-C | 5′-CATGCGGTAGGCGGTATAG | This study |
| ORF311-E | 5′-AATCTCCAAGCGTTTGCTTC | This study |
| OLI-SPEC | 5′-CTCGGTTTTCTGGAAGGCGA | This study |
| THYA-DR-A | 5′-CACAACTCCTGGGGATCAAG | This study |
| THYA-DR-B | 5′-GACCTGCATGTTGTCCCTTC | This study |