TABLE 1.
Bacterial strains, plasmids, and PCR primers
| Strain or plasmid | Genotype or constructiona | Reference or source |
|---|---|---|
| E. coli | ||
| CF1648 | Wild-type MG1655 | 50 |
| CF1652 | As CF1648 but ΔrelA251 | 50 |
| CF1693 | As CF1652 but ΔspoT207 | 50 |
| CF4941 | As 1648 but galK2 zib-563::Tn10 ΔrelA251 | 30 |
| CF4943 | As 4941 but spoT203 | 30 |
| BL21(λDE3) | BL21 (λDE3 lysogen; T7 expression vector) | Novagen |
| Plasmid | ||
| pUC19 | Vector; lacZpo; ColE1 replicon; Apr | |
| pURS1 | As pUC19 but carrying relSeq from S. equisimilis H46A in lacZpo antisense orientation | 30 |
| pET21 | Expression Vector; T7lacpo; pBR replicon; Apr | Novagen |
| pUM99 | As pET21; RelSeq (aa 1-385); C-terminal His tag | This study |
| pBAD18 | Vector; BADpo; pBR replicon; Apr | 17 |
| pBAD22A | Vector; BADpo; pBR replicon; Apr | 17 |
| pUM66 | As pBAD18; RelSeq (aa 1-385); C-terminal His tag | This study |
| pUM108 | As pBAD18; RelSeq (aa 1-224); C-terminal His tag | This study |
| pUM9 | As pBAD18; RelSeq (aa 79-385); C-terminal His tag | This study |
| pUM109 | As pBAD18; RelSeq (aa 1-739); C-terminal His tag | This study |
| pUM110 | As pBAD18; RelSeq (aa 1-385); C-terminal His tag and RelSeq (aa 386-739) coded by separate ORFs | This study |
| pUM111 | As pBAD18; RelSeq (aa 1-385) and RelSeq (aa 386-739) coded by separate ORFs | This study |
| pUM112 | As pBAD18; RelSeq (aa 1-347) and RelSeq (aa 348-739) coded by separate ORFs | This study |
| pUM113 | As pBAD18; RelSeq (aa 386-739 fused to aa 1-385) | This study |
| pUM116 | As pBAD18; SpoT (aa 1-702); C-terminal His tag | This study |
| pUM117 | As pBAD18; SpoT (aa 1-378); C-terminal His tag | This study |
| pLB15 | As pBAD22A; RelA | This study |
| pLB16 | As pBAD22A; RelSeq | This study |
| pLB18 | As pBAD22A; RelA (aa 1-392) fused to RelSeq (aa 383-739) | This study |
| pLB19 | As pBAD22A; RelSeq (aa 1-382) fused to RelA (aa 394-744) | This study |
a
ORF, open reading frame; aa, amino acids.