TABLE 1.

Bacterial strains and plasmids

Strain or plasmidCharacteristic(s)Reference or source
Bacterial strains
    E. coli
        Top 10F mcrA Δ(mrr mutant hsoRMS mutant mcrBC) φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(araA leu)7697 galU galK rpsL endA1 nupGInvitrogen
        XL-1 Blue(F′ proAB laclq ZΔM15 Tn10)supE44 hsdR17 recA1 endA1 gyrA46 thi relA1 lacStratagene
    Y. enterocolitica
        A127/90Clinical isolate from acute gastroenteritis (Japan); serotype O:8, biotype 1B18
        8081Clinical isolate from fatal septicemia (Ohio); serotype O:8, biotype 1B43
        ΔHOPEMTinvInvasin mutant derivative from strain MRS40(pIML421)This work
        8081BF19Integration of the suicide plasmid pBF19 into yspBThis work
        8081FM2Integration of the suicide plasmid pFM2 into ysaNThis work
        8081PT16Integration of the suicide plasmid pPT16 into yspAThis work
        8081PT15Integration of the suicide plasmid pPT15 into yspDThis work
    S. flexneri
        M90TClinical isolate47
        RM97ipgC31
        RM221ipaB32
        SF1068ipaB ipgC28
        RM81ipaC32
Plasmids
    pYV plasmids
        pYV40Wild-type pYV plasmid of Y. enterocolitica E4052
        pABL403pYV40 yopE21 yopHΔ1-352 yopOΔ65-558 yopP23 yopM23, called ΔHOPEM2
        pIML421pYV40 yopE21 yopHΔ1-352 yopP23 yopM23 yopT135 yopOΔ65-558, called ΔHOPEMT19
        pMSK50pYV40 yopE21 yopHΔ1-352 yopOΔ65-558 yopP23 yopM23 yscNΔ169-177, called ΔHOPEMYscNSory and Cornelis, unpublished data
        pAB409pYV40 yopE21 yopHΔ1-352 yopOΔ65-558 yopP23 yopM23 yopBΔ89-217, called ΔHOPEMB2
        pCNK4004pYV40 yopE21 yopHΔ1-352 yopOΔ65-558 yopP23 yopM23 yopBΔ89-217 yopDΔ121-165, called ΔHOPEMBD, obtained by allelic exchange between ΔHOPEMB and pMSL19Neyt and Cornelis, unpublished data
        pCNK4005Introduction of pMSL19 into yopBΔ89-217 mutant bacteria, giving yopBΔ89-217 yopDΔ121-165Neyt and Cornelis, unpublished data
    Expression plasmids and vectors
        pBF15pCNR26 pyopE sycB+ yspB+ yspC+ (from A127/90)This work
        pBF17pBF15 sycB+ yspB(His)6+ yspC+ (A127/90)This work
        pBF20pGEX-6P-1 ptac gst-yspC+ (A127/90)This work
        pBF21pGEX-6P-1 ptac gst-yspB+ (A127/90)This work
        pBF22pGEX-6P-1 ptac gst-sycB+ (A127/90)This work
        pBF23pGEX-6P-1 ptac gst-sycB+ yspB(His)6+ (A127/90)This work
        pBF24pBF23 ptac gst-sycB1-21+ yspB(His)6+ (A127/90)This work
        pBF25pBluescriptSK(−) plac sycB+ (A127/90)This work
        pBF26pBluescriptSK(−) plac sycB+ yspB+ (A127/90)This work
        pBF27pBluescriptSK(−) plac sycB+ yspB+ yspC+ (A127/90)This work
        pBF29pBluescriptSK(−) plac sycB+ yspC+ (A127/90)This work
        pMM100pACYC184 lacl+41
        pPT14pGEX-4T-3 yspD+ (A127/90)This work
        pCNR26Cloning vector with pyopE and optimized ribosome binding site49
        pGEX-4T-3Cloning vector, for generation of fusion with GSTPharmacia Biotech
        pGEX-6P-1Cloning vector, for generation of fusion with GSTPharmacia Biotech
        pBluescript SK(−)Cloning vectorStratagene
    Suicide vectors and mutator plasmids
        pBF19399 bp of yspB (A127/90) cloned into pKNG90AThis work
        pFM2402 bp of ysaN (A127/90) cloned into pKNG90AThis work
        pPT15510 bp of yspD (A127/90) cloned into pPT7This work
        pPT16501 bp of yspA (A127/90) cloned into pPT7This work
        pKNG90ASuicide vector23
        pMS154inv mutator53
        pMSL19pKNG101 yopD+Δ121-26535
        pPT7Derivative of pKNG101 without PstI sites and sacBRThis work