TABLE 1.
Plasmids used in this study
| Plasmids | Relevant characteristics | Reference or source |
|---|---|---|
| pYV plasmids | ||
| pGC153 | pYVe227 yopB-GC153::mini-Mud1 lac ylpA-YL4::Tn3 (does not encode YopB and YopD) | 14 |
| pMRS4071 | pYV40 lcrVΔ3-324 (pYV40 mutated with pMRS71) | This work |
| pPW2269 | pYVe227 sycD mutant (out-of-frame deletion between nucleotides 111 and 255 of sycD; encodes a 43-residue truncated SycD) | 53 |
| pYV40 | Wild-type PYV plasmid from Y. enterocolitica E40 | 49 |
| pYVe227 | Wild-type pYV plasmid from Y. enterocolitica W227 | 9 |
| Clones and vectors | ||
| pBC18R | 10 | |
| pBC19R | 10 | |
| pCNR21 | pBC19R PyopE yopE | This work |
| pCNR26 | pBC19R PyopE SD T7 NdeI yopE | This work |
| pCN29 | pGEX-KG yopD (encodes GST and YopD) | This work |
| pCN40 | pGEX-KG yopB sycD (encodes GST, YopB and SycD) | This work |
| pCNG42 | pGEX-KG gst-lcrV yopB sycD (encodes GST-LcrV, YopB and SycD) | This work |
| pCNG50 | pGEX-KG gst-lcrV sycD (encodes GST-LcrV and SycD) | This work |
| pGEX-KG | ptac gst oripBR322 | 20 |
| pMRS20 | pBluescriptII SK+ and PCR-amplified fragment (by using MIPA271 and MIPA64) containing lcrRGV | 45 |
| pMRS28 | pBC18R and EcoRI fragment from pMRS20 (encoding lcrRGV under own promoter) | This work |
| pMRS44 | pT7-7 and PCR-amplified lcrG (by using MIPA354 and MIPA355) from pMRS20 cloned into the NdeI-SalI site | This work |
| pMRS46 | pGEX-KG gst-lcrV (encodes GST-LcrV) | 45 |
| pMRS50 | pGEX-KG gst-lcrG yopD (encodes GST-LcrG and YopD) | 45 |
| pMRS52 | pMRS20 lcrVΔ224-266 (created by site-directed mutagenesis with MIPA352) | This work |
| pMRS56 | pMRS20 lcrVΔ2-32 (created by site-directed mutagenesis with MIPA356) | This work |
| pMRS64 | pBluescript II SK+ and PCR-amplified fragment (by using MIPA384 and MIPA289) from pYV40 containing lcrG, lcrV, and sycD genes | This work |
| pMRS65 | pMRS64, NruI site just before codon 3 of lcrV | This work |
| pMRS67 | pMRS65, SmaI site just after codon 324 of lcrV | This work |
| pMRS69 | pMRS67 lcrVΔ3-324 | This work |
| pMRS70 | pMRS101, SalI-XbaI fragment of pMRS69 | This work |
| pMRS72 | pCNR26 and PCR-amplified fragment (by using MIPA354 and MIPA122) from pYV40 containing lcrG, lcrV, and sycD genes | This work |
| pMRS74 | pMSL56 with cyaA replaced by a PCR-amplified fragment (by using MIPA121 and MIPA317) from pYV40 containing sycD, yopB, and yopD genes | This work |
| pMRS75 | pMRS46 lcrG (encodes GST-LcrV and LcrG) | This work |
| pMRS78 | pMRS75 lcrVΔ224-266 (encodes GST-LcrVΔ224-266 and LcrG) | This work |
| pMRS83 | pMRS75 lcrVΔ2-32(encodes GST-LcrVΔ2-32 and LcrG) | This work |
| pMRS84 | pMRS50 lcrV (lcrG replaced by lcrV) (encodes GST-LcrV and YopD) | This work |
| pMSL56 | pTM100 PyopE cyaA | M.-P. Sory |
| pPW64 | pBC18R sycD | 53 |
| Suicide vector and mutator | ||
| pMRS101 | oriR6KoriE1 sacBR oriTRK2strAB | 44 |
| pMRS71 | pMRS70 ΔoriE1 (encoding lcrVΔ3-324) | This work |