TABLE 1.
Bacterial strains and plasmids used in this work
| Strain or plasmid | Relevant phenotype or genotype | Source or reference |
|---|---|---|
| R. sphaeroides | ||
| 2.4.1 | Wild type | 31 |
| PRRB1 | 2.4.1 derivative, Δ prrB::ΩSpr/Str | 8 |
| PRRA2 | 2.4.1 derivative, Δ prrA::ΩSpr/Str | 8 |
| E. coli | ||
| DH5α | (Φ80dlacZΔM15)ΔlacU169 recA1 endA1 hsdR17 supE44 thil gyrA96 relA1 | 13 |
| S17-1 | Pro− Res− Mob+recA; integrated plasmid RP4-Tc::Mu-Km::Tn7 | 29 |
| Plasmids | ||
| pUC19 | Apr; lacPOZ′ | 32 |
| pBSIIKS+ | Apr; lacPOZ′ | Stratagene |
| pT7-7 | Apr; T7 promoter, RBS, and translation start codon overlapping with NdeI site | 30 |
| pRK415 | Tcr; Mob+lacZα IncP | 15 |
| pU1655 | pBSIIKS+::0.47-kb StyI fragment of pufBA | J. K. Lee |
| pUCP6.37 | pUC19::3.2-kb PvuII fragment containing the rrnB operon | S. C. Dryden |
| pUI1643 | pBSIIKS+::4.0-kb BamHI-HindIII fragment containing prrB and prrA | 8 |
| pUI1649 | pRK415/BamHI-Asp718, 1.9-kb SmaI-ClaI fragment containing prrB; prrB is in divergent orientation to lacZ | 8 |
| pUI1663 | Spr Str Kmr; IncQ; puf::lacZYA′ | 6 |
| pCF1010 | Spr Str Tcr; IncQ | 18 |
| pPRRB1-1 | pBSIIKS+::2.0-kb ApaI fragment containing prrB | This study |
| pPRRB2 | pBSIIKS+::429-bp PCR product containing six His codons at the 3′ end of prrB; prrB is in divergent orientation to lacZ | This study |
| pPRRB3 | pPRRB1-1::400-bp PflMI-KpnI from pPRRB2 | This study |
| pPRRB4 | pRK415::1.65-kb PstI-KpnI fragment containing prrB from pPRRB3; prrB is in colinear orientation to lacZ | This study |
| pPRRB5 | pUC19::1.65-kb PstI-KpnI fragment containing prrB from pPRRB3 | This study |
| pBLP1,3,5 | pBSIIKS+::1.65-kb PstI-KpnI fragment containing prrB with five-Ala codon insertion mutation and six His codons at the 3′ end of prrB | This study |
| pLP1-pLP5 | pPRRB4 in which five Ala codons are inserted in prrB | This study |
| pTM1Δ | pPRRB4 with a deletion mutation in prrB | This study |
| pOYH1 | pT7-7::1.42-kb NdeI-PstI fragment containing prrB | This study |
| pOYH2 | pBSIIKS+::1.43-kb XbaI-PstI fragment from pOYH1 | This study |
| pA-9 | pRK415::1.45-kb XbaI-KpnI fragment from pOYH2; prrB is in colinear orientation to lacZ | This study |
| pPRRBLAC | pCF1010::0.6-kb PstI-XbaI fragment of prrB promoter region | This study |