TABLE 1.
Strains and plasmids used in this work
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Strains | ||
| R. sphaeroides | ||
| APP11 | ΔappA::Tpr | 7 |
| APS1 | APP11 ppsR::ΩKmr | 10 |
| PS3 | APP11 ppsR3 (L107-P) | 10 |
| PS8 | APP11 ppsR8 (L345-P) | 10 |
| PS18 | APP11 ppsR18 (C251-R) | 10 |
| PS23 | APP11 ppsR23 (H90-R) | 10 |
| PS33 | APP11 ppsR33 (L248-P) | 10 |
| PS36 | APP11 ppsR36 (K422-E) | 10 |
| PS40 | APP11 ppsR40 (C424-R)a | 10 |
| PS41 | APP11 ppsR41 (L403-P) | 10 |
| PS48 | APP11 ppsR48 (L28-P) | This work |
| PS50 | APP11 ppsR50 (L249-P) | This work |
| PS52 | APP11 ppsR52 (L27-P) | This work |
| PS58 | APP11 ppsR58 (R242-C) | This work |
| PS66 | APP11 ppsR66 (N176-I) | This work |
| PS69 | APP11 ppsR69 (G321-V) | This work |
| PS72 | APP11 ppsR72 (G117-E) | This work |
| PS77 | APP11 ppsR77 (G288-E) | This work |
| PS80 | APP11 ppsR80 (R339-Q) | This work |
| P. denitrificans ATCC 17741 | Wild type | American Type Culture Collection |
| E. coli | ||
| S17-1 | Tra+ donor for conjugations | 26 |
| DH5α phe | Plasmid maintenance | 5 |
| JM109 | Overexpression of the GST-PpsR protein | Lab collection |
| Plasmids | ||
| pCF400Δ | Smr/Spr Kmr IncQ, puc::lacZ | 9 |
| pLX200 | Smr/Spr IncQ, bchF::lacZ | 9 |
| pLX14b | Smr/Spr IncQ, bchF::lacZ | 10 |
| pLX14P | Smr/Spr IncQ, bchF::lacZ, ppsR | 10 |
| pRK415 | Tcr IncP, vector | 14 |
| pPNs | pRK415::ppsR (wild type) | 9 |
| pPS#c | pRK415::ppsR# | This work |
| pPSΔ10 | pRK415::ppsRΔ10 (deletion of residues 2–10 of PpsR) | This work |
| pPSΔ40 | pRK415::ppsRΔ40 (deletion of residues 2–39 of PpsR) | This work |
| pPSΔ59-117 | pRK415::ppsRΔ59-117 (deletion of residues 60–116 of PpsR) | This work |
| pP[R] | pRK415::ppsRΔ406-464 (deletion of residues 406–464 of PpsR) | 9 |
| pBg[Ns] | pRK415::ppsRΔ1-356 (deletion of residues 1–356 of PpsR) | 9 |
| pGEX-2TK | Vector for overexpression GST fusions | Amersham Pharmacia Biotech |
| pGpps | pGEX-2TK::ppsR | This work |
| pRK2013 | Helper plasmid for conjugations | 6 |
a
This mutation was erroneously described as C424-A in reference 10, although proper nucleotide substitution was presented.
b
Plasmid pLX14, similar to pLX200, contains a bchF::lacZ transcriptional fusion. The difference between these two plasmids is in the point of fusion, which does not affect bchF expression (9, 10).
c
Symbol # refers to the number of the ppsR mutation.