Table 1.
Strains and Plasmids used in this study.
| Name | Genotype/Description | Origin/Reference |
|---|---|---|
| BACTERIAL STRAINS | ||
| Escherichia coli DH5α | F−Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (, ) phoA supE44 λ– thi-1 gyrA96 relA1 | Invitrogen |
| E. coli BL21 (DE3) | F−ompT hsdSB() gal dcm (DE3) pRARE | Novagen |
| E. coli JM110 | F− traD36 lacIqΔ(lacZ)M15proA+B+/rpsL(StrR) thr leu thi laY galK galT ara fhuA dam dcm glnV44 Δ(lac-proAB) | ATCC |
| EEPrbP | BL21 (DE3) carrying p15TV-prbP. Ampr | Gardner et al., 2016 |
| EEPrbP-N107A | BL21 (DE3) carrying p15TV-prbP(N107A). Ampr | This work |
| EEPrbP-G109A | BL21 (DE3) carrying p15TV-prbP(G109A). Ampr | This work |
| EEPrbP-I114A | BL21 (DE3) carrying p15TV-prbP(I114A). Ampr | This work |
| EEPrbP-R144A | BL21 (DE3) carrying p15TV-prbP(R144A). Ampr | This work |
| EEPrbP-R147A | BL21 (DE3) carrying p15TV-prbP(R147A). Ampr | This work |
| EEPrbP-E148A | BL21 (DE3) carrying p15TV-prbP(E148A). Ampr | This work |
| EERNAPD | BL21 (DE3) carrying p15TV-rpoD, pACYCDuet-rpoB-rpoC, pRSFDuet-rpoZ-rpoA. Ampr. Cmr. Kanr | This work |
| ECPRPLK | DH5α carrying pMiniT-PrplK. Ampr | This work |
| PLASMIDS | ||
| p15TV-L | Expression vector for purification of proteins by nickel affinity chromatography. Ampr | Cheryl Arrowsmith, (Addgene plasmid # 26093) |
| p15TV-prbP | prbP gene from L. asiaticus cloned in the BseRI site of p15TV-L. Ampr | Gardner et al., 2016 |
| p15TV-prbP(N107A) | p15TV-prbP with mutation for N107A. Ampr | This work |
| p15TV-prbP(G109A) | p15TV-prbP with mutation for G109A. Ampr | This work |
| p15TV-prbP(I114A) | p15TV-prbP with mutation for I114A. Ampr | This work |
| p15TV-prbP(R144A) | p15TV-prbP with mutation for R144A. Ampr | This work |
| p15TV-prbP(R147A) | p15TV-prbP with mutation for R147A. Ampr | This work |
| p15TV-prbP(E148A) | p15TV-prbP with mutation for E148A. Ampr | This work |
| p15TV-rpoD | rpoD gene from L. asiaticus cloned in the NdeI-BamHI site of p15TV-L. Ampr | This work |
| pACYCDuet-1 | Expression vector for co-purification of protein complexes by nickel affinity chromatography. Cmr | Novagen |
| pRSFDuet-1 | Expression vector for co-purification of protein complexes by nickel affinity chromatography. Kanr | Novagen |
| pACYCDuet-rpoB | rpoB gene from L. asiaticus cloned in the FseI-PvuI site of pACYCDuet-1 plasmid. Cmr | This work |
| pRSFDuet-rpoZ | rpoZ gene from L. asiaticus cloned in the SalI-NotI site of pRSFDuet-1 plasmid. Kanr | This work |
| pACYCDuet-rpoB-rpoC | rpoC and rpoB genes from L. asiaticus cloned in the PstI-NotI and FseI-PvuI sites of pACYCDuet-1 plasmid respectively. Cmr | This work |
| pRSFDuet-rpoZ-rpoA | rpoA and rpoZ genes from L. asiaticus cloned in the EcoRV-KpnI and SalI-NotI sites of pRSFDuet-1 plasmid respectively. Kanr | This work |
| pMiniT | Cloning vector for PCR amplicon | NEB |
| pMiniT-PrplK | Promoter region (−230 to +54 bp) of rplK gene cloned into insertion site of pMiniT. Ampr | This work |