TABLE 1.
Plasmids and Synechocystis sp. strain PCC 6803 mutants used and constructed in this study (see Fig. 1)
| Plasmid or mutant | Descriptiona |
|---|---|
| Plasmids | |
| pGG | pGEM7 containing ggpS (sll1566) as a 2.1-kb BamHI fragment obtained by PCR with ggpS5 and ggpS3 primers; final size, 5.2 kb |
| pGΔG::Km2 | pGG containing an inactivated ggpS gene (aphII gene is inserted in the SnaBI/StuI deletion in the opposite direction of ggpS); final size, 6.1 kb |
| pGΔG::Sm2 | pGG containing an inactivated ggpS gene (smr gene is inserted in the SnaBI/StuI deletion in the opposite direction of ggpS); final size, 6.7 kb |
| pGD | pGEM7 containing glpD (sll1085) as a 1.7-kb EcoRI/XhoI fragment obtained by PCR with glpD5 and glpD3 primers; final size, 4.7 kb |
| pGD::Km2 | pGD containing an inactivated glpD gene (aphII gene is inserted at the unique BlpI site in the opposite direction of glpD); final size, 5.9 kb |
| pGD::Sm2 | pGD containing an inactivated glpD gene (smr gene is inserted at the unique BlpI site in the opposite direction of glpD); final size, 6.7 kb |
| pGK | pGEM7 containing glpK (slr1672) as a 1.5-kb EcoRI/BamHI fragment obtained by PCR with glpK5 and glpK3 primers; final size, 4.5 kb |
| pGK::Km2 | pGK containing an inactivated glpK gene (aphII gene is inserted at the BstEII deletion in the opposite direction of glpK); final size, 5.6 kb |
| pBADGGPS | pBAD/HisB containing ggpS (sll1566) as a 1.6-kb BglII/KpnI fragment obtained by PCR with ggpSE5′ and ggpSE3′ used for ggpS overexpression in E. coli LMG194; final size, 5.7 kb |
| Mutants | |
| ΔGK2 | Salt-sensitive Synechocystis mutant impaired in ggpS (sll1566) obtained after transformation of the WT with pGΔG::Km2 |
| ΔGS2 | Salt-sensitive Synechocystis mutant impaired in ggpS (sll1566) obtained after transformation of the WT with pGΔG::Sm2 |
| DK2 | Salt-tolerant Synechocystis mutant impaired in glpD (sll1085) obtained after transformation of the WT with pGD::Km2 |
| ΔKK2 | Salt-tolerant Synechocystis mutant impaired in glpK (slr1672) obtained after transformation of the WT with pGK::Km2 |
| DS2ΔKK2 | Salt-tolerant Synechocystis mutant impaired in glpD (sll1085) and glpK (slr1672) obtained after transformation of ΔKK2 with pGD::Sm2 |
a
Salt tolerance of the mutants was tested by the ability to grow in 0.55 M NaCl or to lyse at this salt concentration.