TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant property(ies) | Source or reference |
|---|---|---|
| E. coli strains | ||
| Novablue | Recipient for cloning experiments | Novagen |
| BL21(DE3) | Host for overexpression from the T7 promoter | Novagen |
| RP6894 | ΔmotAB | J. S. Parkinson |
| Salmonella sp. strain SJW1103 | Wild type for motility and chemotaxis | 46 |
| Plasmids | ||
| pTrc99A | Cloning vector | Pharmacia |
| pHMK11 | pTrc expression vector | This study |
| pACTrc | pTrc promoter, p15A replication origin, lacIq Cmr | G. M. Fraser |
| pET19b | T7 expression vector | Novagen |
| pET22b | T7 expression vector | Novagen |
| pHMK1609 | pHMK11/MotA+MotB-His8 | This study |
| pNSK6 | pET19b/MotBC-His6 | This study |
| pNSK7 | pHMK11/PelBL::MotBC-His6a | This study |
| pNSK8 | pTrc99A/MotBC-His6 | This study |
| pNSK11 | pET19b/MotBC | This study |
| pNSK28 | pHMK11/PelBL::MotBCa | This study |
| pNSK31 | pACTrc/MotA+MotB | This study |
| pNSK32 | pACTrc/MotA+MotB-His8 | This study |
| pNSK6-R218W | pET19b/MotBC(R218W)-His6 | This study |
| pNSK6-Δ(197-210) | pET19b/MotBC(Δ197-210)-His6 | This study |
| pNSK6-T197I/R218W | pET19b/MotBC(T197I/R218W)-His6 | This study |
| pNSK7-T197I | pHMK11/PelBL::MotBC(T197I)-His6a | This study |
| pNSK7-D198N | pHMK11/PelBL::MotBC(D198N)-His6a | This study |
| pNSK7-S215F | pHMK11/PelBL::MotBC(S215F)-His6a | This study |
| pNSK7-R218W | pHMK11/PelBL::MotBC(R218W)-His6a | This study |
| pNSK7-R223H | pHMK11/PelBL::MotBC(R223H)-His6a | This study |
| pNSK7-Δ(197-210) | pHMK11/PelBL::MotBC(Δ197-210)-His6a | This study |
| pNSK7-Δ(211-226) | pHMK11/PelBL::MotBC(Δ211-226)-His6a | This study |
| pNSK7-T197I/R218W | pHMK11/PelBL::MotBC(T197I/R218W)-His6a | This study |
a
A MotB fragment coding for residue 78 to C-terminal residue 309 (MotBC) was fused to a PelB leader sequence derived from pET22b at the N terminus and fused to His6 at the C terminus.