Table 1.

Plasmids used in this study

PlasmidsRelevant characteristicsReference or source
pYV plasmids
 pGC153pYVe227yopB-GC153::mini-Mud1 lac ylpA-YL4::Tn3 (does not encode YopB and YopD) 14
 pMRS4071pYV40 lcrV Δ3-324 (pYV40 mutated with pMRS71)This work
 pPW2269pYVe227sycD mutant (out-of-frame deletion between nucleotides 111 and 255 of sycD; encodes a 43-residue truncated SycD) 53
 pYV40Wild-type PYV plasmid from Y. enterocoliticaE40 49
 pYVe227Wild-type pYV plasmid fromY. enterocolitica W227 9
Clones and vectors
 pBC18R 10
 pBC19R 10
 pCNR21pBC19R PyopE yopE This work
 pCNR26pBC19R PyopE SD T7 NdeI yopE This work
 pCN29pGEX-KG yopD (encodes GST and YopD)This work
 pCN40pGEX-KG yopB sycD (encodes GST, YopB and SycD)This work
 pCNG42pGEX-KG gst-lcrV yopB sycD (encodes GST-LcrV, YopB and SycD)This work
 pCNG50pGEX-KG gst-lcrV sycD (encodes GST-LcrV and SycD)This work
 pGEX-KGptac gst ori pBR322 20
 pMRS20pBluescriptII SK+ and PCR-amplified fragment (by using MIPA271 and MIPA64) containinglcrRGV 45
 pMRS28pBC18R andEcoRI fragment from pMRS20 (encoding lcrRGV under own promoter)This work
 pMRS44pT7-7 and PCR-amplifiedlcrG (by using MIPA354 and MIPA355) from pMRS20 cloned into the NdeI-SalI siteThis work
 pMRS46pGEX-KG gst-lcrV (encodes GST-LcrV) 45
 pMRS50pGEX-KGgst-lcrG yopD (encodes GST-LcrG and YopD) 45
 pMRS52pMRS20lcrV Δ224-266 (created by site-directed mutagenesis with MIPA352)This work
 pMRS56pMRS20lcrV Δ2-32 (created by site-directed mutagenesis with MIPA356)This work
 pMRS64pBluescript II SK+ and PCR-amplified fragment (by using MIPA384 and MIPA289) from pYV40 containing lcrG, lcrV, andsycD genesThis work
 pMRS65pMRS64,NruI site just before codon 3 of lcrV This work
 pMRS67pMRS65, SmaI site just after codon 324 oflcrV This work
 pMRS69pMRS67lcrV Δ3-324 This work
 pMRS70pMRS101,SalI-XbaI fragment of pMRS69This work
 pMRS72pCNR26 and PCR-amplified fragment (by using MIPA354 and MIPA122) from pYV40 containing lcrG, lcrV, andsycD genesThis work
 pMRS74pMSL56 withcyaA replaced by a PCR-amplified fragment (by using MIPA121 and MIPA317) from pYV40 containing sycD, yopB, and yopD genesThis work
 pMRS75pMRS46lcrG (encodes GST-LcrV and LcrG)This work
 pMRS78pMRS75 lcrV Δ224-266 (encodes GST-LcrVΔ224-266 and LcrG)This work
 pMRS83pMRS75 lcrV Δ2-32(encodes GST-LcrVΔ2-32 and LcrG)This work
 pMRS84pMRS50 lcrV (lcrG replaced bylcrV) (encodes GST-LcrV and YopD)This work
 pMSL56pTM100 PyopE cyaA M.-P. Sory
 pPW64pBC18R sycD 53
Suicide vector and mutator
 pMRS101 ori R6K ori E1 sacBR oriT RK2 strAB 44
 pMRS71pMRS70 Δori E1 (encodinglcrV Δ3-324)This work