TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Reference or source |
|---|---|---|
| Strains | ||
| 86-24 | Wild-type EHEC strain (serotype O157:H7) | 23a |
| DH5α | supE44 lacU169 (80 lacZ M15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Stratagene |
| VS94 | 86-24 luxS mutant | 71 |
| BH68 | 86-24 qseD mutant | This study |
| BH85 | BH68 complemented with pBH22 | This study |
| BH86 | BH68 complemented with pBH23 | This study |
| BW25113 (K-12) | rrnB3 ΔlacZ4787 hsdR514 Δ(araBAD)567 Δ(rhaBAD)568 rph-1 | 13 |
| D7 | BW25113 qseD mutant; Keio collection; yjiE7 | 5a |
| BH51 | D7 complemented with plasmid pBH22 | This study |
| BH52 | D7 complemented with plasmid pBH23 | This study |
| D166 | Michigan State E. coli strain TW05550 | NIAID STEC Center |
| TB226A | Michigan State E. coli strain TW04257 | NIAID STEC Center |
| B2F1 | Michigan State E. coli strain TW07506 | NIAID STEC Center |
| VP08 | Michigan State E. coli strain TW04584 | NIAID STEC Center |
| DEC 16A | Michigan State E. coli strain TW02918 | NIAID STEC Center |
| 78/92 | Michigan State E. coli strain TW05608 | NIAID STEC Center |
| BH294-10 | Michigan State E. coli strain TW02898 | NIAID STEC Center |
| BH262C-8 | Michigan State E. coli strain TW02897 | NIAID STEC Center |
| D55 | Michigan State E. coli strain TW04549 | NIAID STEC Center |
| VP12 | Michigan State E. coli strain TW04588 | NIAID STEC Center |
| 97-3250 | Michigan State E. coli strain TW07814 | NIAID STEC Center |
| 493/89 | Michigan State E. coli strain TW06555 | NIAID STEC Center |
| DEC 5B | Michigan State E. coli strain TW02719 | NIAID STEC Center |
| DEC 5A | Michigan State E. coli strain TW00587 | NIAID STEC Center |
| DEC 5C | Michigan State E. coli strain TW01959 | NIAID STEC Center |
| DEC 5D | Michigan State E. coli strain TW00947 | NIAID STEC Center |
| DEC 5E | Michigan State E. coli strain TW00962 | NIAID STEC Center |
| 5905 | Michigan State E. coli strain TW05353 | NIAID STEC Center |
| G5101 | Michigan State E. coli strain TW05356 | NIAID STEC Center |
| 413 | Michigan State E. coli strain TW05359 | NIAID STEC Center |
| EDL931 | Michigan State E. coli strain TW02303 | NIAID STEC Center |
| EDL932 | Michigan State E. coli strain TW02299 | NIAID STEC Center |
| EDL933 | Michigan State E. coli strain TW02302 | NIAID STEC Center |
| L40 | Yeast reporter strain; MATα trp1 leu2 his3 LYS2::lexA-HIS3 URA3::lexA-lacZ | 79 |
| B2HPC | L40 transformed with pLX-YopJ plus pVP-MEKK (yeast 2-hybrid positive control) | 53a |
| B2HNC | L40 transformed with pLX-Laminin plus pVP16 (yeast 2-hybrid negative control) | 53a |
| BH129 | L40 transformed with pVPSA plus pLXSA | This study |
| BH130 | L40 transformed with pVPSA plus pLXLA | This study |
| BH131 | L40 transformed with pVPSA plus pLXSD | This study |
| BH132 | L40 transformed with pVPSA plus pLXLD | This study |
| BH133 | L40 transformed with pVPLA plus pLXSA | This study |
| BH134 | L40 transformed with pVPLA plus pLXLA | This study |
| BH135 | L40 transformed with pVPLA plus pLXSD | This study |
| BH136 | L40 transformed with pVPLA plus pLXLD | This study |
| BH137 | L40 transformed with pVPSD plus pLXSA | This study |
| BH138 | L40 transformed with pVPSD plus pLXLA | This study |
| BH139 | L40 transformed with pVPSD plus pLXSD | This study |
| BH140 | L40 transformed with pVPSD plus pLXLD | This study |
| BH141 | L40 transformed with pVPLD plus pLXSA | This study |
| BH142 | L40 transformed with pVPLD plus pLXLA | This study |
| BH143 | L40 transformed with pVPLD plus pLXSD | This study |
| BH144 | L40 transformed with pVPLD plus pLXLD | This study |
| BH157 | L40 transformed with pVPSL plus pLXSL | This study |
| BH158 | L40 transformed with pVPSL plus pLXLL | This study |
| BH159 | L40 transformed with pVPSL plus pLXSD | This study |
| BH160 | L40 transformed with pVPSL plus pLXLD | This study |
| BH161 | L40 transformed with pVPLL plus pLXSL | This study |
| BH162 | L40 transformed with pVPLL plus pLXLL | This study |
| BH163 | L40 transformed with pVPLL plus pLXSD | This study |
| BH164 | L40 transformed with pVPLL plus pLXLD | This study |
| BH165 | L40 transformed with pVPSD plus pLXSL | This study |
| BH166 | L40 transformed with pVPSD plus pLXLL | This study |
| BH167 | L40 transformed with pVPLD plus pLXSL | This study |
| BH168 | L40 transformed with pVPLD plus pLXLL | This study |
| BH29 | BH68 complemented with pBH24 | This study |
| BH31 | BH68 complemented with pBH26 | This study |
| Plasmids | ||
| pACYC184 | Cloning vector | NEB |
| pBAD-MycHIS (A) | C-terminal Myc-His expression vector | Invitrogen |
| pBH22 | K-12 qseD region inserted into Tet of pACYC184 | This study |
| pBH23 | 8624 qseD region inserted into Tet of pACYC184 | This study |
| pKD46 | λ Red helper plasmid | 13 |
| pKD3 | λ Red template plasmid | 13 |
| pCP20 | λ Red resolvase plasmid | 13 |
| pVP16 | Yeast library plasmid | 79 |
| pLex-ADE | Yeast bait plasmid; pBTM116 with the insertion of ADE2 at the PvuII site | 79 |
| pLX-YopJ | pLEX-ADE expressing YopJ | 53a |
| pVP-MEKK | pVP-16 expressing MEKK | 53a |
| pLX-Laminin | pLEX-ADE expressing laminin | 53a |
| pVPSA | pVP16 expressing “short” QseA | This study |
| pVPLA | pVP16 expressing “long” QseA | This study |
| pVPSD | pVP16 expressing “short” QseD | This study |
| pVPLD | pVP16 expressing “long” QseD | This study |
| pVPSL | pVP16 expressing “short” LrhA | This study |
| pVPLL | pVP16 expressing “long” LrhA | This study |
| pLXSA | pLEX-ADE expressing “short” QseA | This study |
| pLXLA | pLEX-ADE expressing “long” QseA | This study |
| pLXSD | pLEX-ADE expressing “short” QseD | This study |
| pLXLD | pLEX-ADE expressing “long” QseD | This study |
| pLXSL | pLEX-ADE expressing “short” LrhA | This study |
| pLXLL | pLEX-ADE expressing “long” LrhA | This study |
| pET28 | N-terminal His-tagged T7 expression construct | Novagen |
| pBH24 | K-12 QseD in pBAD-MycHIS (A) | This study |
| pBH26 | 86-24 QseD in pBAD-MycHIS (A) | This study |
| pMK208 | QseA-His (N-terminal) in pET28 | This study |