TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Genotype description, or sequence (5′→3′)a | Source or referenceb |
|---|---|---|
| Strains | ||
| S. enterica serovar Typhimurium | ||
| TH6238 | motA5461::MudJ | |
| TH8059 | motA5461::MudJ fliA6088 (A. aeolicus GTG-fliA+) | |
| TH8120 | motA5461::MudJ flgM6095 (A. aeolicus flgM+) | |
| TH8122 | motA5461::MudJ fliA6088 flgM6095 | |
| TH8194 | motA5461::MudJ ΔflgM5794::FRTc | |
| TH8196 | motA5461::MudJ fliA6088 ΔflgM5794::FRT | |
| TH8197 | motA5461::MudJ ΔflgM5794::FRT ΔfliA5805::tetRA | |
| TH4098 | fliC5050::MudJ fljBe,n,xvh2 | |
| TH8061 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 (A. aeolicus GTG-fliA+) | |
| TH8123 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 | |
| TH8125 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6095 | |
| TH8198 | fliC5050::MudJ fljBe,n,xvh2 ΔflgM5794::FRT | |
| TH8200 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 ΔflgM5794::FRT | |
| TH8201 | fliC5050::MudJ fljBe,n,xvh2 ΔflgM5794::FRT ΔfliA5805::tetRA | |
| MST4190 | leuBCD485 trp::[Spcr PlacT7-RNAP lacI] | S. Maloy |
| TH9031 | leuBCD485 trp::[Spcr PlacT7-RNAP lacI] motA5461::MudJ fliA6325 (C. trachomatis L2 GTG-fliA+) ΔflgM5794::FRT/pJK627 | |
| TH9032 | leuBCD485 trp::[Spcr PlacT7-RNAP lacI] motA5461::MudJ fliA6325 ΔflgM5794::FRT/pET15b | |
| TH9033 | leuBCD485 trp::[Spcr PlacT7-RNAP lacI] motA5461::MudJ fliA6325/pET15b | |
| TH9040 | leuBCD485 trp::[Spcr PlacT7-RNAP lacI] fliC5050::MudJ fljBe,n,xvh2 fliA6325 ΔflgM5794::FRT/pJK627 | |
| TH9041 | leuBCD485 trp::[Spcr PlacT7-RNAP lacI] fliC5050::MudJ fljBe,n,xvh2 fliA6325 ΔflgM5794::FRT/pET15b | |
| TH9042 | leuBCD485 trp::[Spcr PlacT7-RNAP lacI] fliC5050::MudJ fljBe,n,xvh2 fliA6325/pET15b | |
| TH8604 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6237 (A. aeolicus flgM A→G −6 bp from ATG) | |
| TH8605 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6238 (A. aeolicus flgM A→C −5 bp from ATG) | |
| TH8606 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6239 (A. aeolicus flgM A→G −4 bp from ATG) | |
| TH8607 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6240 (A. aeolicus flgM ATG→GTG M1V) | |
| TH8608 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6241 (A. aeolicus flgM AAG→AGG K19R) | |
| TH8609 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6242 (A.aeolicus flgM Δ bp +55-56) | |
| TH8610 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6243 (A. aeolicus flgM ΔA bp +177 from ATG) | |
| TH8611 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6244 (A. aeolicus flgM AAA→AAG K64K, ΔA bp +199 from ATG) | |
| TH8612 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6245 (A. aeolicus fliA CCT→TCT P4S) | |
| TH8613 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6246 (A. aeolicus fliA ACA→TCA T27S) | |
| TH8614 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6247 (A. aeolicus fliA ATA→ATG 129M) | |
| TH8615 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6248 (A. aeolicus fliA AGA→GGA R149G) | |
| TH8616 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6249 (A. aeolicus fliA ACG→ATG 1175M) | |
| TH8617 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6250 (A. aeolicus fliA GAA→GGA E189G) | |
| TH8618 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6251 (A. aeolicus fliA GAA→GGA E199G) | |
| TH9607 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6245 ΔflgM5794::FRT | |
| TH9608 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6246 ΔflgM5794::FRT | |
| TH9609 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6247 ΔflgM5794::FRT | |
| TH9610 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6248 ΔflgM5794::FRT | |
| TH9611 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6249 ΔflgM5794::FRT | |
| TH9612 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6250 ΔflgM5794::FRT | |
| TH9647 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 fliA6251 ΔflgM5794::FRT | |
| TH3467 | proAB47/F′128 (pro-lac) zzf-3833::Tn10dTc[del-25] (T-POP) | 34 |
| TH4387 | fliA5059::Tn10dTc fljBe,n,xvh2 | |
| TH7953 | flgM6085::tetRA/pKD46 | |
| TH8009 | ΔfliA5805::tetRA/pKD46 | |
| TH8010 | ΔfliA5805::tetRA motA5461::MudJ/pKD46 | |
| TH8011 | ΔfliA5805::tetRA fliC5050::MudJ fljBe,n,xvh2/pKD46 | |
| TH8349 | fliC5050::MudJ fljBe,n,xvh2 fliA6090 flgM6085::tetRA/pKD46 | |
| TH8350 | fliC5050::MudJ fljBe,n,xvh2 flgM6095 ΔfliA5805::tetRA/pKD46 | |
| TH8142 | fliA6088 (A. aeolicus GTG-fliA+) | |
| TH8982 | fliA6325 (C. trachomatis L2 GTG fliA+) | |
| TH9065 | ΔfliA5647::FRT/pBAD24 | |
| TH9066 | ΔfliA5647::FRT/pMC147 | |
| TH9067 | ΔfliA5647::FRT/pJK558 | |
| TH9068 | ΔfliA5647::FRT/pJK629 | |
| E. coli | ||
| DH5α | φ80dlacZΔM15 endA1 hsdR17 (rK−mK−) supE44 thi-1 recA1 gyrA (Nalr (lacZYA-argF) U169) | New England Biolabs |
| Plasmids | ||
| pMC147 | (pBAD24-fliA S. enterica serovar Typhimurium) | |
| pJK558 | (pBAD24-fliA A. aeolicus) | |
| pJK629 | (pBAD24-rspD C. trachomatis) | |
| pJK627 | (pET15b-rsbW C. trachomatis L2) | |
| pKD46 | (pBAD- λ-Red (γ β exo) | 8 |
| pBAD24 | (Para cloning vector pBR322 ori, Apr) | 14 |
| pET15b | Novagen | |
| pMS531 | (S. enterica serovar Typhimurium fliA) | 42 |
| pLF28 | (PBAD-rspD C. trachomatis) | 39 |
| pET28a-rsbW | C. trachomatis L2 | T. P. Hatch |
| Primers | ||
| FliAdeltetR | CTCATTTAACGCAGGGCTGTTTATCGTGAATATAGGTCGCGCATGATCGCACCC GAAAAGTttaagacccactttcaca | |
| FliAdeltetA | CTTTTCGGGTGCGATCATGCGCGACCTATATTCACGATAAACAGCCCTGCGTTA AATGAGTctaagcacttgtctcctg | |
| FlgMAUGtetR | AGCTGGCCGCTACAACGTAACCCTCGATGAGGATAAATAAttaagacccactttcacatt | |
| FlgMAUGtetA | TGCTAACGGGTTTCAAAGGTGAGGTACGGTCAATGCTCATctaagcacttgtctcctg | |
| AAGTG | TAATCATGCCGATAACTCATTTAACGCAGGGCTGTTTATCgtgaaaaacccttacagcaacc | |
| AA28R | CGTTGTGCGGCACTTTTCGGGTGCGATCATGCGCGACCTAtagaggattagagagcatttc | |
| AAFLGM1 | AGCTGGCCGCTACAACGTAACCCTCGATGAGGATAAATAAatggtgaacaggattcaactc | |
| AAFLGM2 | CATCTGGTCAAGTATTTCTGACAAACGAGTCATACGCTTAcgtaaaaaactctatcagtcc | |
| CTFLIAGTG | TAATCATGCCGATAACTCATTTAACGCAGGGCTGTTTATCgtgaagactcacgatctcgc | |
| CTFLIASTOP | CGTTGTGCGGCACTTTTCGGGTGCGATCATGCGCGACCTAaagcagactggacaatgtac | |
| AQAA1 | AAGAATTCATGAAAAACCCTTACAGCAACC | |
| AQAA2 | ACAAAGCTTATAGAGGATTAGAGAGCATTTC | |
| FliA#3 | GGCGCTACAGGTTACATAAG | |
| FliA#4 | TAGTCTATACGTTGTGCGGC | |
| FlgM5′UP | GAACCGTCGATTCTGATG | |
| FlgN reverse | GTCTTCAGGTCATTCAG |
a
In primer sequences, lowercase indicates homology to the tetRA element.
b
All strains were constructed for this study unless otherwise noted.
c
FRT, FLP recognition target scar (8).