TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Description/genome/sequence (5′-3′) | Reference or source |
|---|---|---|
| E. coli strains | ||
| DH5α | F− endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 hsdR17(rK− mK+) λ− | Invitrogen |
| BL21 | F− ompT gal dcm lon hsdSB(rB− mB−) λ (DE3)pLysS (Cmr)a | New England Biolabs |
| W3110 | rph-1 IN(rrnD-rrnE) | Coli Genetic Stock Centre |
| EJR02 | W3110 ΔzapD | This work |
| Plasmids | ||
| pEJR005b | pET28A with WT ftsZ from E. coli | 27 |
| pEJR031 | pBAD24 with WT His6-zapD from E. coli | This work |
| pEJR031_R16A | pBAD24 with R16A His6-zapD from E. coli | This work |
| pEJR031_R20A | pBAD24 with R20A His6-zapD from E. coli | This work |
| pEJR031_R56A | pBAD24 with R56A His6-zapD from E. coli | This work |
| pEJR031_E114A | pBAD24 with E114A His6-zapD from E. coli | This work |
| pEJR031_R116A | pBAD24 with R116A His6-zapD from E. coli | This work |
| pEJR031_R124A | pBAD24 with R124A His6-zapD from E. coli | This work |
| pEJR031_R176A | pBAD24 with R176A His6-zapD from E. coli | This work |
| pEJR031_R221A | pBAD24 with R221A His6-zapD from E. coli | This work |
| pEJR031_R225A | pBAD24 with R225A His6-zapD from E. coli | This work |
| Primers for mutagenesis | ||
| R16A_F | CAATGCGCAGCCATGTAGCCATTTTTTCATTTAGTGGATGTTCA | This work |
| R16A_R | TGAACATCCACTAAATGAAAAAATGGCTACATGGCTGCGCATTG | This work |
| R20A_F | TGCTGAATCAAAAACTCAATGGCCAGCCATGTACGCATTTTTTC | This work |
| R20A_R | GAAAAAATGCGTACATGGCTGGCCATTGAGTTTTTGATTCAGCA | This work |
| R56A_F | GTGCGGACTTCGCCGGCCTCGAAAACATCCAG | This work |
| R56A_R | CTGGATGTTTTCGAGGCCGGCGAAGTCCGCAC | This work |
| E114A_F | GAGCAATCAAACGATCTGCACGCAGAAATTGCCCG | This work |
| E114A_R | CGGGCAATTTCTGCGTGCAGATCGTTTGATTGCTC | This work |
| R116A_F | ACGCACCAGAGCAATCAAAGCATCTTCACGCAGAAATTGC | This work |
| R116A_R | GCAATTTCTGCGTGAAGATGCTTTGATTGCTCTGGTGCGT | This work |
| R124A_F | CTGGGATGCTCAGTGCCTGACGCACCAGAG | This work |
| R124A_R | CTCTGGTGCGTCAGGCACTGAGCATCCCAG | This work |
| R176A_F | GGGCCGACTGGGCAATTAAATCCAGCACCATGGT | This work |
| R176A_R | ACCATGGTGCTGGATTTAATTGCCCAGTCGGCCC | This work |
| R221A_F | GGCATAAAACGAATGGCAAAAGCGCTCTTATGTCCGGAAATTTG | This work |
| R221A_R | CAAATTTCCGGACATAAGAGCGCTTTTGCCATTCGTTTTATGCC | This work |
| R225A_F | GTGTCCAGCGGCATAAAAGCAATGGCAAAACGGCTCTT | This work |
| R225A_R | AAGAGCCGTTTTGCCATTGCTTTTATGCCGCTGGACAC | This work |
a
Cmr, chloramphenicol resistance.