Table 1.
Characteristics of the plasmids used in this study
| Plasmid | Relevant feature(s)a | Reference or source |
|---|---|---|
| Analysis of protein-protein interactions with the bacterial two-hybrid system | ||
| pUT18 | Ampr plasmid encoding the T18 domain of the Bordetella pertussis adenylate cyclase in frame with an upstream multiple-cloning site | 24 |
| pKT25 | Kmr plasmid encoding the T25 domain of the B. pertussis adenylate cyclase in frame with a downstream multiple-cloning site | 24 |
| pUT18-zip | pUT18 with the CS for the self-interacting Zip control protein | 24 |
| pKT25-zip | pKT25 with the CS for the self-interacting Zip control protein | 24 |
| pUT18-merA | pUT18 with the full-length Synechocystis merA CS (slr1849) | This study |
| pKT25-merA | pKT25 with the full-length Synechocystis merA CS | This study |
| pUT18-arsC | pUT18 with the full-length Synechocystis arsC CS (slr0946) | This study |
| pKT25-arsC | pKT25 with the full-length Synechocystis arsC CS | This study |
| pUT18-grx1 | pUT18 with the full-length Synechocystis grx1 CS (slr1562) | 3 |
| pKT25-grx1 | pKT25 with the full-length Synechocystis grx1 CS | 3 |
| pUT18-grx2 | pUT18 with the full-length Synechocystis grx2 CS (ssr2061) | 3 |
| pKT25-grx2 | pKT25 with the full-length Synechocystis grx2 CS | 3 |
| pKT18-grx3 | pKT25 with the full-length Synechocystis grx3 CS (slr1846) | This study |
| pKT25-grx3 | pKT25 with the full-length Synechocystis grx3 CS | This study |
| Gene inactivation in Synechocystis | ||
| pGEMT | Ampr AT overhang cloning vector | Promega |
| pUC4K | Source of the Kmr marker gene | Pharmacia |
| pHP45Ω | Source of the Smr Spr marker gene | 39 |
| pΔgrx1::Kmr | Kmr cassette for deletion of the grx1 CS | 3 |
| pΔgrx2::Smr/Spr | Smr Spr for deletion of the grx2 CS | 3 |
| pmerA | pGEMT with the 534-bp merA CS preceded by its 177-bp upstream region | This study |
| pΔmerA::Kmr | pmerA with the Kmr marker in a SmaI site created at 84 bp of MerA CS | This study |
| Gene expression in Synechocystis | ||
| pFC1 | Replicating plasmid for heat-inducible gene expression in Synechocystis | 32 |
| pFmerA | pFC1 with the merA CS cloned between NdeI and EcoRI for heat-inducible production of MerA | This study |
| pSB2A | Replicating plasmid for expressing genes from their own promoter | 34 |
| pSmerA | pSB2A expressing the merA gene from its 150-bp promoter region | This study |
| pSmerAC78S | pSmerA where cysteine 78 of MerA is replaced by a serine | This study |
| pSgrx1 | pSB2A expressing the grx1 gene from its 150-bp promoter region | This study |
| pSgrx1C86S | pSgrx1 where cysteine 86 of Grx1 is replaced by a serine | This study |
| Protein production in E. coli | ||
| pET21b(+) | Plasmid for fusion of the 6×His tag at the C terminus of proteins | Novagen |
| pETM30 | Plasmid for fusion of the GST tag at the N terminus of proteins | Novagen |
| pTrc2 | Plasmid for fusion of the cMyc and 6×His tags at the C terminus of proteins | Invitrogen |
| pgrx1-6His | pET21b(+) with the grx1 CS cloned at the NdeI-EcoRI sites | This study |
| pgrx1-6HisC31S | pgrx1-6His with the C31S mutation in grx1 | This study |
| pgrx1-6HisC86S | pgrx1-6His with the C86S mutation in grx1 | This study |
| pmerA-6His | pTrc2 with the merA CS cloned at the BamHI-EcoRI sites | This study |
| pmerAC78S-6His | pmerA-6His harboring the C78S mutation in merA | This study |
| pgrx2-6His | pTRc2 with the grx2 CS cloned at the BamHI-EcoRI sites | This study |
| pGST-grx1 | pETM30 with the grx1 CS cloned at the SalI-XhoI sites | This study |
| pMaL-c2 | Plasmid for fusion of the MBP tag at the C terminus of proteins | New England BioLabs |
| pSS9 | pMaL-c2 with the merA CS cloned at the EcoRI-BamHI sites | This study |
a
CS, coding sequence.