TABLE 1.
E. coli strains and plasmids used in this work
| Strain, plasmid | Genotype or description | Ref. or source |
|---|---|---|
| E. coli strains | ||
| BL21(DE3) | hsdS (rB−mB−) gal dcm ompT λ(DE3), T7 RNA polymerase gene under control of the lacUV5 promoter | 51 |
| C41 | Derivative of BL21 (DE3) | 52 |
| SIP1332 | C41 pSP130/29 | This study |
| SIP1453 | As SIP1332 but tonB | This study |
| SIP1461 | As SIP1332 but fhuA | This study |
| Plasmids | ||
| pHM10 | pACYC184 encoding FyuA | 53 |
| pSP130/29 | pET25b encoding FyuA with His tag at N terminus of mature protein | This study |
| pSP130/33 | pET22b encoding Pim-His6 | This study |
| pSP130/64 | pET22b encoding Pst(1–167)-His6 (PstRT) | This study |
| pSP130/66 | pET22b encoding Pst(164–357)-His6 (PstA) | This study |
| pSP130/70 | pET22b encoding Pst(1–357)His6 (Pst) | This study |
| pSP130/73 | pET22b encoding PstQ301A-His6 | This study |
| pSP130/88 | pET22b encoding PstP187A-His6 | This study |
| pSP130/92 | pET22b encoding PstT201A-His6 | This study |
| pSP130/110 | pET22b encoding PstE178A-His6 | This study |
| pSP130/467 | pET22b encoding PstS89C/S285C-His6 | This study |
| pSP130/480 | pET22b encoding PstD338A-His6 | This study |
| pSP130/486 | pET22b encoding PstD207A-His6 | This study |
| pSP130/258 | pET22b encoding PstΔ10–15-His6 | This study |
| pSP130/262 | pET22b encoding PstΔ10–20-His6 | This study |
| pSP130/375 | pET22b encoding T4L-His6; T4L refers to LYS_BPT4 Swiss-Prot P00720 with replacement of the two intrinsic cysteine residues (C221T/C264A) | This study |
| pSP130/187 | pET24d encoding Pst-T4L-His6 | This study |
| pSP130/432 | pET24d encoding Pst-T4LD294C/R321C-His6 | This study |
| pSP130/379 | pET24d encoding Pst-T4LI176C/R321C-His6 | This study |
| pSP130/441 | pET24d encoding Pst-T4LI176C/L331C-His6 | This study |
| pSP130/452 | pET24d encoding Pst-T4LS257C/Q289C-His6 | This study |
| pSP130/151 | Encodes MalE (1–26)-Pst with a C-terminal His tag (MalE′-Pst) controlled by the araBAD promoter, araC, Ampr | This study |
| pSP130/152 | Encodes MalE (1–26)-Pst (164–357) with a C-terminal His tag (MalE′-PstA) under control of the araBAD promoter, araC, Ampr | This study |
| pSP130/176 | Encodes MalE′-PstE178A with a C-terminal His tag under control of the araBAD promoter, araC, Ampr | This study |
| pSP130/178 | Encodes MalE′-PstA (E178A) with a C-terminal His tag under control of the araBAD promoter, araC, Ampr | This study |