TABLE 2.
Plasmids, vectors, and reagents used in this study
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Strain, strain background (Salmonella enterica serovar Typhimurium) | SJW1103 | (30) | Wild type | This strain can be obtained from the Fraser lab upon request. |
| Strain, strain background (Salmonella enterica serovar Typhimurium) | recA null | (7) | recA gene replaced with kanamycin resistance cassette | This strain can be obtained from the Fraser lab upon request. |
| Strain, strain background (Salmonella enterica serovar Typhimurium) | flgD null | (17) | flgD gene replaced with kanamycin resistance cassette | This strain can be obtained from the Fraser lab upon request. |
| Strain, strain background (Salmonella enterica serovar Typhimurium) | flgE null | This work | flgE gene replaced with kanamycin resistance cassette | This strain can be obtained from the Fraser lab upon request. |
| Strain, strain background (Salmonella enterica serovar Typhimurium) | flhB P28T, fliHI null, flgM null | (14) | P28T mutation introduced into chromosomal flhB gene, flgHI genes removed by scarless mutagenesis, flgM gene replaced by spectinomycin resistance cassette | This strain can be obtained from the Fraser lab upon request. |
| Strain, strain background (Salmonella enterica serovar Typhimurium) | flhB P28T, fliHI null, flgM null, fliMN null | This work | P28T mutation introduced into chromosomal flhB gene, flgHI and fliMN genes removed by scarless mutagenesis, flgM gene replaced by spectinomycin resistance cassette | This strain can be obtained from the Fraser lab upon request. |
| Strain, strain background (Salmonella enterica serovar Typhimurium) | flgE 234(3FLAG)235 | This work | Triple flag tag inserted between residue 234 and 234 of flgE gene | This strain can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE | (7) | 1–234, FLAG × 3, 235–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆Ct | This work | 1–234, FLAG × 3, 235–359 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgG | (7) | 1–144, FLAG × 3, 145–260 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgG∆Ct | This work | 1–144, FLAG × 3, 145–217 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgC | This work | 1–69, FLAG × 3, 70–134 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgC∆Ct | This work | 1–69, FLAG × 3, 70–90 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgD | This work | 1–172, FLAG × 3, 173–232 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgD∆Ct | This work | 1–172, FLAG × 3, 173–190 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆341–350 | This work | 1–234, FLAG × 3, 235–340, 351–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆351–360 | This work | 1–234, FLAG × 3, 235–360, 361–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆361–370 | This work | 1–234, FLAG × 3, 235–360, 371–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆371–380 | This work | 1–234, FLAG × 3, 235–370, 381–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆381–390 | This work | 1–234, FLAG × 3 , 235–380, 391–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆391–403 | This work | 1–234, FLAG × 3, 235–390 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE V366A | This work | 1–234, FLAG × 3, 235–403 aa, V366A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE E371A | This work | 1–234, FLAG × 3, 235–403 aa, E371A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE L372A | This work | 1–234, FLAG × 3, 235–403 aa, L372A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE V373A | This work | 1–234, FLAG × 3, 235–403 aa, V373A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE I376A | This work | 1–234, FLAG × 3, 235–403 aa, I376A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE V377A | This work | 1–234, FLAG × 3, 235–403 aa, V377A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE Q379A | This work | 1–234, FLAG × 3, 235–403 aa, Q379A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE R380A | This work | 1–234, FLAG × 3, 235–403 aa, R380A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE Y382A | This work | 1–234, FLAGx3, 235–403 aa, Y382A | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgEshort + linker | (7) | 1–8, 4× (Gly-Ser-Thr-Asn-Ala-Ser), 33–234, FLAG × 3, 235–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgEshort | (7) | 1–8, 33–234, FLAG × 3, 235–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Recombinant DNA reagent | pTrc99a FlgE∆GRM | This work | 1–39, 44–234, FLAG × 3, 235–403 aa | This vector can be obtained from the Fraser lab upon request. |
| Antibody | Anti-FLAG (mouse monoclonal) | Sigma-Aldrich | Cat # F3165, RRID:AB_259529 | Mouse monoclonal against FLAG tag (1:1,000) |
| Antibody | Anti-FlgD (rabbit polyclonal) | (17) | Rabbit polyclonal against Salmonella FlgD (1:1,000) This antibody can be obtained from the Fraser lab upon request. | |
| Antibody | Anti-FliC (rabbit polyclonal) | (17) | Rabbit polyclonal against Salmonella FliC (1:1,000). This antibody can be obtained from the Fraser lab upon request. | |
| Antibody | Anti-FlgL (rabbit polyclonal) | (7) | Rabbit polyclonal against Salmonella FlgL (1:1,000). This antibody can be obtained from the Fraser lab upon request. | |
| Antibody | Anti-FlgN (rabbit polyclonal) | (15) | Rabbit polyclonal against Salmonella FlgN (1:1,000). This antibody can be obtained from the Fraser lab upon request. | |
| Antibody | Anti-FlhA (rabbit polyclonal) | (15) | Rabbit polyclonal against Salmonella FlhA (1:1,000). This antibody can be obtained from the Fraser lab upon request. | |
| Antibody | Anti-GroEL (rabbit polyclonal) | Sigma-Aldrich | Cat # G6532 | Rabbit polyclonal against FLAG tag (1:40,000) |