TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Escherichia coli strains | ||
| NovaBlue | Recipient for cloning experiments | Novagen |
| BL21(DE3)/pLysS | Overproduction of proteins | Novagen |
| HMS174(DE3)/pLysS | Overproduction of proteins | Novagen |
| Salmonella strains | ||
| JR501 | For converting E. coli plasmids to Salmonella compatibility | 19 |
| KK1013 | Deletion of region III | 9 |
| MKM11 | ΔfliH | 6 |
| MKM11gK | ΔfliH flgK::Tn10; defective in region I | This study |
| MKM11hB | ΔfliH flhB::Tn10; defective in region II | This study |
| MMA10xx series | ΔfliH flhA pseudorevertants from MKM11 | This study |
| MMA11xx series | flhA second-site mutants from corresponding MMA10xx strain | This study |
| MMB10xx series | ΔfliH flhB pseudorevertants from MKM11 | This study |
| MMB11xx series | flhB second-site mutants from corresponding MMB10xx strain | This study |
| SJW1103 | Wild-type strain for motility and chemotaxis | 25 |
| SJW2936 | Deletion of region II | 22 |
| SJW1429 | fliH | 15 |
| Plasmids | ||
| pET19b | Vector | Novagen |
| pTrc99A | Vector | Pharmacia |
| pMM310 | pET19b/His-FliHa | 17 |
| pMM306 | pTrc99A/His-FliH | 17 |
| pMM405 | pET19b/His-FliJa | 17 |
| pMM406 | pTrc99A/His-FliJ | 17 |
| pMM1701 | pET19b/His-FliIa | 17 |
| pMM1702 | pTrc99A/His-FliI | 17 |
| pMM1704 | pTrc99A/His-FliI(R7C/L12P) | 16 |
| pMM1706 | pTrc99A/His-FliI(K188I) | 16 |
| pMM1708 | pTrc99A/His-FliI(Y363S) | 16 |
| pBGtI(Δ1-7) | pTrc99A/His-FliI(Δ1-7)b | This study |
| pBGtICAT | pTrc99A/His-FliICAT | This study |
| pIFF700 | pTrc99A/His-FLAG-FliI | 4 |
| pIFF7001, etc. | pTrc99A/His-FLAG-FliIΔ1, etc. | Fan Fan, unpublished |
a
Although in many cases we made pET19b-based versions of plasmids, we include in the table only those encoding the wild-type alleles of FliH, FliI, and FliJ.
b
FliI(Δ1-7) was obtained in a previous study (18) as a clostripain proteolysis fragment with an apparent mass of 48 kDa and was given the name FliICL48 in that paper. The N-His-tagged version of FliICL48 in the present paper has been given the more descriptive name His-FliI(Δ1-7).