Table 1.
Bacterial strains and genetic constructs used in this study.
| Strain | Relevant characteristics of virulence plasmid | References |
|---|---|---|
| Y. enterocolitica strains | ||
| MRS40 | Wild-type pYV plasmid of Y. enterocolitica E40 (pYVe40) ΔblaA | Sory et al., 1995 |
| IML421asd (ΔHOPEMTasd) | pYVmrs40 yopOΔ2−427 yopE21 yopHΔ1−352 yopM23 yopP23 yopT135 Δasd | Kudryashev et al., 2013 |
| AD4051 | pYVmrs40 ΔsctD | Diepold et al., 2010 |
| AD4085 | pYViml421asd egfp-sctQ | Kudryashev et al., 2013 |
| AD4306 | pYViml421asd egfp-sctD | Diepold et al., 2015 |
| AD4483 | pYViml421asd egfp-sctL ΔsctD | Diepold et al., 2017 |
| ADTM4521 | pYViml421asd mCherry-sctL | This study |
| IM41 | pYVmrs40 sctW45 (does not encode sctW = yopN) | Boland et al., 1996 |
| CH4006 | pYViml421asd sctFS5C | This study |
| Plasmid | Relevant characteristics | References |
| Plasmids | ||
| pACYC184 | Expression vector | New England Biolabs |
| pKNG101 | oriR6K sacBR+ oriTRK2 strAB+ (suicide vector) | Kaniga et al., 1991 |
| pAD626 | pACYC184::yopH1−17-bla (β-lactamase from pBAD::His-B cloned in-frame with first 17 amino acids of Y. enterocolitica yopH) | This study |
| pAD627 | pACYC184::bla | This study |
| pADTM521 | pKNG101 mCh-sctL (pamCherry and flexible linker cloned in-frame at the N-terminus of sctL) | Diepold et al., 2017 |
| pCH06 | pKNG101 sctFS5C (serine to cysteine point mutation of amino acid 5 in sctF) | This study |
Strains and constructs used in this study. Y. enterocolitica strains are based on either the wild-type strain MRS40 or the multi-effector knock-out strain ΔHOPEMTasd (IML421asd), which lacks the virulence effectors YopH, YopO, YopP, YopE, YopM, and YopT, and is auxotrophic for diaminopimelic acid as result from a mutation in the aspartate-β-semialdehyde (asd) gene. The knock-out of the virulence effectors and the asd mutation is used for biosafety purposes, allowing these strains to be handled in a biosafety 1 level environment.