Table 3.
Bacterial strains and plasmids used in the study.
| Strain/plasmid | Genotype/description | Source/reference |
|---|---|---|
| STRAINS | ||
| E. coli Top10 | General cloning strain | Invitrogen |
| E. coli BL21 (DE3) | Strain for protein expression | Invitrogen |
| E. coli J53 (RP4) | Helper strain for tri-parental mating | Wolk et al., 1984 |
| Synechocystis sp. PCC6803 | Wild type strain | Stanier et al., 1971 |
| ΔPII | glnB− strain of Synechocystis sp. PCC6803 | Hisbergues et al., 1999 |
| Complementation | ΔPII strain complemented with PII-Venus | This study |
| Complementation S49E | ΔPII strain complemented with PIIS49E-Venus | This study |
| PLASMIDS | ||
| pET15b | Expression vector for His-tagged proteins | Novagen |
| pET15accB | Expression of Synechocystis His-BCCP | This study |
| pCY216 | Expression of E. coli BirA | Chapman-Smith et al., 1994 |
| pET16baccAD | Expression of E. coli His-AccA and AccD | Soriano et al., 2006 |
| pET16baccC | Expression of E. coli His-AccC | Soriano et al., 2006 |
| pTRPETBCCPn | Expression of E. coli His-BCCP | Rodrigues et al., 2014 |
| pASK-IBA3 | Expression vector for Strep-taged proteins | IBA life sciences |
| pASK-IBA3glnB | Expression of C-terminally tagged GlnB from Synechococcus elongatus. PCC7942 | Heinrich et al., 2004 |
| pASK-IBA3glnBS49D | Synechococcus GlnB variant S49D | Espinosa et al., 2006 |
| pASK-IBA3glnBS49E | Synechococcus GlnB variant S49E | Heinrich et al., 2004 |
| pASK-IBA3glnBR45A | Synechococcus GlnB variant R45A | This study |
| pASK-IBA3glnBR47A | Synechococcus GlnB variant R47A | This study |
| pASK-IBA3glnBS49G | Synechococcus GlnB variant S49G | This study |
| pASK-IBA3glnBY51A | Synechococcus GlnB variant Y51A | This study |
| pASK-IBA3glnBT52A | Synechococcus GlnB variant T52A | This study |
| pASK-IBA3glnBE54A | Synechococcus GlnB variant E54A | This study |
| pASK-IBA3glnBE85A | Synechococcus GlnB variant E85A | This study |
| pASK-IBA3glnBI86N | Synechococcus GlnB variant I86N | Fokina et al., 2010b |
| pASK-IBA3glnBR103H | Synechococcus GlnB variant R103H | This study |
| pASK-IBA3glnBSc | Expression of C-terminally tagged GlnB from Synechocystis sp. PCC6803 | This study |
| pASK-IBA3glnBS49CSc | Synechocystis GlnB variant S49C | This study |
| pASK-IBA3glnBS49DSc | Synechocystis GlnB variant S49D | This study |
| pASK-IBA3glnBS49ESc | Synechocystis GlnB variant S49E | This study |
| pVZ322 | Broad host range expression vector | Grigorieva and Shestakov, 1982 |
| pVZ322-PII-Ven | Expression of wild type GlnB with the fluorophore Venus at the C-terminus | This study |
| pVZ322-PIIS49E-Ven | Expression of GlnB S49E variant with the fluorophore Venus at the C-terminus | This study |