TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Property(ies) | Source or reference |
|---|---|---|
| Strains | ||
| E. coli K-12 | ||
| TOP10 F′ | F′ [lacIq Tn10(Tetr)]mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| Y. enterocolitica | ||
| W22703 | O:9 serotype, pYVe227 | 8a |
| CT133 | pYVe227 sycD [SycDΔ7-49]a | 2 |
| VTL2 | pYVe227 yopD [YopDΔ9-306] | 27a |
| Y. pseudotuberculosis | ||
| YPIII pIB102 | pIB yadA::Tn5 | 6a |
| YPIII pIB68 | pIB yscS [YscSΔ19-59] yadA::Tn5 | 5a |
| Plasmids and constructs | ||
| pTrcHisB | Apr; expression vector for creation of N-terminal His6-containing fusions | Invitrogen |
| pFlag-CTC | Apr; expression vector for creation of C-terminal Flag-tagged fusions | Sigma |
| pEF/myc/cyto | Neor; eukaryotic expression vector used as source of neomycin phosphotransferase (npt) | Invitrogen |
| pHT-LcrH | lcrH PCR amplified from Y. pseudotuberculosis YPIII pIB102 (nt 4-504) in pTrcHisB fused in frame with the His6-containing leader sequence | This study |
| pSycD-FT | PCR-amplified Y. enterocolitica sycD (nt 4-504) in pFlag-CTC fused in frame with C-terminal Flag epitope tag | This study |
| pScc1-FT | PCR-amplified scc1 (CT088; nt 4-438)b in pFlag-CTC fused in frame with C-terminal Flag epitope tag | This study |
| pScc2-FT | PCR-amplified scc2 (CT576; nt 4-696) in pFlag-CTC fused in frame with C-terminal Flag epitope tag | This study |
| pScc3-FT | PCR-amplified scc3 (CT862; nt 4-594) in Flag-CTC fused in frame with C-terminal Flag epitope tag | This study |
| pCopB2NT | PCR-amplified copB2 N terminus (CT861; nt 4-951) in pFlag-CTC in frame with C-terminal Flag epitope tag | This study |
| pCopBnpt | pFlag-CTC expressing a chimeric product consisting of CopB (aa 1-120)c, Npt, and a C-terminal Flag epitope tag | This study |
a
Numbers in brackets correspond to residues deleted from the gene product.
b
The C. trachomatis genome designation and the respective nucleotides (nt) amplified are provided.
c
Numbers refer to residues of the C. trachomatis product present in the respective chimeric protein. aa, amino acids.