TABLE 1.
Strains and plasmids used in this study
| Species and strain or plasmid | Description | Reference or source |
|---|---|---|
| C. trachomatis ATCC VR-346 | Serovar F/IC-Cal-13 | ATCC |
| E. coli | ||
| KS1 | MC1000 F′ lacIq integrated with a single copy of promoter plac OR2-62 controlled by lacZ in the chromosome | 8 |
| ZY101 | MC1000 F′ lacIq integrated with a single copy of Plac_hctB-35(TAAAGTTT)::lacZ on an F′episome | This study |
| ZY102 | MC1000 F′ lacIq integrated with a single copy of Plac_hctB-35(gAAAGTTT)::lacZ on an F′ episome | This study |
| BN317 | MC1000 F′ lacIq integrated with a single copy of placCons-35C::lacZ on an F′ episome | 34 |
| S. enterica serovar Typhimurium | ||
| TH7034 | Isogenic with wild-type strain LT2; fliA5886 [R91C L207P] fliC5050::MudJ | K. T. Hughes |
| TH5504 | Isogenic with wild-type strain LT2; fliA5647::FRT | K. T. Hughes |
| Plasmids | ||
| pFW11 | 45 | |
| pS28H | ParaB-His6 chlamydial fliA | 39 |
| pES28H | ParaB-His6E. coli fliA | 39 |
| pHσ28Ct-Ec4 | ParaB-His6 hybrid fliA; encoding σ28Ct residues 1-178 fused to region 4 of σ28Ec residues 166 to 239 | This study |
| pHσ28Ec-Ct4 | ParaB-His6 hybrid fliA; encoding σ28Ec (residues 1-165) fused to region 4 of σ28Ct (residues 179-253) | This study |
| pRV5 | Promoter probe vector containing less lacZ; ligation of the XmnI-SphI fragment of pQF50KgroE and the MfeI (blunted)-SphI fragment of pTara; origin of replication of p15A; Cmr | This study |
| pRVPhctB | pRV5 with promoter of chlamydial hctB (−125 to +45) | This study |
| pQF50KgroE | Promoter reporter vector containing PgroE::lacZ | 24 |
| pTara | Expression vector carrying ParaB-T7RNAP | 47 |
| pACλcI | PlacUV5-directed synthesis of the λCI protein | 8 |
| pACλcI-β-flap | PlacUV5-directed synthesis of the λCI protein fused via three alanines to residues 858-946 of the β subunit of E. coli RNAP | 25 |
| pACλcI-β-flapCt | PlacUV5-directed synthesis of the λCI protein fused via three alanines to residues 798-887 of the β subunit of C. trachomatis RNAP | This study |
| pBRα | PlacUV5/Plpp-directed synthesis of the α subunit of E. coli RNAP | 8 |
| pBRαLN | PlacUV5/Plpp-directed synthesis of N terminus (residues 1-248) of the α subunit of E. coli RNAP | 8, 10 |
| pBRα-σ66 | PlacUV5/Plpp-directed synthesis of the αNTD fused to C. trachomatis σ66 region 4 (residues 468-571 of σ66) | This work |
| pBRα-σ70 | PlacUV5/Plpp-directed synthesis of the αNTD fused to E. coli σ70 region 4 (residues 528-613 of σ70) | 10 |
| pBRα-σ70D581G | PlacUV5/Plpp-directed synthesis of the αNTD fused to E. coli σ70 region 4 carrying the D581G substitution. | 25 |
| pBRα-σ28 | PlacUV5/Plpp-directed synthesis of the αNTD fused to chlamydial σ28Ct region 4 (residues 148-253 of σ28) | This study |
| pBRα-σ28E206G | pBRα-σ28 encoding the E206G substitution in the β moiety of the fusion | This study |
| pBRα-σ28Y214C | pBRα-σ28 encoding the Y214C substitution in the β moiety of the fusion | This study |
| pBRα-σ28E222G | pBRα-σ28 encoding the E222G substitution in the β moiety of the fusion | This study |
| pBRα-σ28Q236L | pBRα-σ28 encoding the Q236L substitution in the β moiety of the fusion | This study |
| pBRα-σ28L243K | pBRα-σ28 encoding the L243K substitution in the β moiety of the fusion | This study |
| pGPhctB | Contains a chlamydial-hctB-controlled guanine-less cassette and an E. coli fliC promoter-controlled guanine-less cassette; Apr | 40 |
| pGPhctBΔup | Derivative of pGPhctB with the AT-rich sequence upstream of the promoter −35 element deleted | 40 |
| pGS9 | Derivative of pGPhctB containing a 9-bp spacer between the −35/−10 elements | 40 |
| pGS14 | Derivative of pGPhctB containing a 14-bp spacer between the −35/−10 elements | 40 |
| pGPtar | Contains an E. coli tar-controlled guanine-less cassette and an E. coli fliC promoter-controlled guanine-less cassette; Apr | 40 |
| pGPtar+ | Derivative of pGPtar fusing a DNA sequence upstream of PhctB to the core promoter of Ptar | 40 |
| pGHR | Contains a PhctB-controlled guanine-less cassette and a chlamydial rRNA P1 promoter-controlled guanine-less cassette | This study |
| pLF28 | Low-copy-number expression vector carrying chlamydial fliA; Tcr | 39 |
| pLF28a | Derivative of pLF28 with a silent mutation at nucleotide position 723 to remove the natural HindIII site | This study |