TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| Strains | ||
| E. coli K24 | EHEC O157, stx1 | This study |
| E. coli EDL933 | EHEC O157, stx1, stx2 | 24 |
| E. coli 86-24 | EHEC O157, stx2 | 10 |
| E. coli TS26 | EHEC O26H11, stx1 | This study |
| E. coli K47 | EHEC O157, stx2 but did not detect Stx2 | This study |
| E. coli BL21 | F−ompT hsdSB(rB− mB−) dcm gal | Promega |
| E. coli DH5αλpir | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 deoR (lacZYA-argF)U169 λpir+ | 6 |
| E. coli S17-1λpir | pro res mod+RP4-2 Tet::Mu-Kan::Tn7 | 27 |
| E. coli EDLN | Spontaneous variant of EDL933; Nalr | This study |
| E. coli N20-8 | Isogenic mutant of EDLN; stxB1 (N32S) | This study |
| E. coli N1-12 | Isogenic mutant of EDLN; stxB2 (S31N) | This study |
| Plasmids | ||
| pTrcHis2A | Ptrc laca; Codons for 6 histidine residues, Ampr | Invitrogen |
| pWM91 | Suicide vector; Ampr SacB+ | 18 |
| pi-1 | stxB1 in pTrcHis2A | This study |
| piii-1 | stxB2 in pTrcHis2A | This study |
| piS-3 | stxB1 His6 in pTrcHis2A | This study |
| piiiS-1 | stxB2 His6 in pTrcHis2A | This study |
| pAH-5 | (RBR-StxB1)a-stxB2 fusion His6 in pTrcHis2A | This study |
| pCH-6 | stxB110EstxB2 fusion His6 in pTrcHis2A | This study |
| pDH-22 | stxB123KstxB2 fusion His6 in pTrcHis2A | This study |
| pEH-7 | stxB135NstxB234NstxB1 fusion His6 in pTrcHis2A | This study |
| pOH-1 | stxB123KstxB2 fusion His6 in pTrcHis2A | This study |
| pFH-5 | stxB148MstxB2 fusion His6 in pTrcHis2A | This study |
| pHH-8 | (RBR-StxB1)a-stxB29EstxB1 fusion His6 in pTrcHis2A | This study |
| pNH-3 | stxB110EstxB222KstxB1 fusion His6 in pTrcHis2A | This study |
| pPH-2 | stxB135NstxB247MstxB1 fusion His6 in pTrcHis2A | This study |
| pXH-7 | stxB123KstxB226KstxB1 fusion His6 in pTrcHis2A | This study |
| pYH-2 | stxB127KstxB234NstxB1 fusion His6 in pTrcHis2A | This study |
| pL29Y-1 | stxB1(L29Y) His6 in pTrcHis2A | This study |
| pF30W-1 | stxB1(F30W) His6 in pTrcHis2A | This study |
| pN32S-1 | stxB1(N32S) His6 in pTrcHis2A | This study |
| pY28L-1 | (RBR-StxB1)a-stxB2(Y28L) His6 in pTrcHis2A | This study |
| pW29F-2 | (RBR-StxB1)a-stxB2(W29F) His6 in pTrcHis2A | This study |
| pS31N-6 | (RBR-StxB1)a-stxB2(S31N) His6 in pTrcHis2A | This study |
| pStx1-4 | stx1 in pTrcHis2A | This study |
| p2AKH-2 | stx2 in pTrcHis2A | This study |
| pStx1N32S-1 | stx1 mutant (B subunit, N32S) in pTrcHis2A | This study |
| pStx2S31N-22 | stx2 mutant (B subunit, N32S) in pTrcHis2A | This study |
| pWN32S-20 | stx1 mutant (B subunit, N32S) in pWM91 | This study |
| pWS31N-2 | stx2 mutant (B subunit, N32S) in pWM91 | This study |
a
RBR-StxB1 means ribosomal binding region of StxB1.