TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Reference or source |
|---|---|---|
| Strains | ||
| DH5α | Vector propagation, recA1 endA1 | |
| BL21(DE3)pLysS | BL21 with IPTG-inducible T7 polymerase | 90 |
| TW14359 | Wild-type strain from the 2006 outbreak, western USA | 91 |
| EcRJM-1 | TW14359 ΔescN | 20 |
| EcRJM-6 | TW14359 ΔrcsB | 20 |
| EcRJM-10 | TW14359 rcsB-FLAG | 20 |
| EcRJM-11 | TW14359 ΔgrvA | 20 |
| EcRJM-12 | TW14359 ΔrcsB ΔgrvA | 20 |
| EcRJM-13 | TW14359 tir-FLAG | 20 |
| EcRJM-35 | TW14359 ΔgrlR::kan | 20 |
| EcRJM-72 | TW14359 ΔrcsB/pRJM-20 | 20 |
| EcRJM-73 | TW14359 ΔlacZ | This study |
| EcRJM-74 | TW14359/pRJM-23 | This study |
| EcRJM-75 | TW14359/pRJM-24 | This study |
| EcRJM-76 | TW14359/pRJM-25 | This study |
| EcRJM-77 | TW14359/pRJM-26 | This study |
| EcRJM-78 | TW14359 ΔgrvA/pRJM-23 | This study |
| EcRJM-79 | TW14359 ΔgrvA/pRJM-24 | This study |
| EcRJM-80 | TW14359 ΔgrvA/pRJM-25 | This study |
| EcRJM-81 | TW14359 ΔgrvA/pRJM-26 | This study |
| EcRJM-82 | TW14359 ΔgrvA ΔgadE::kan/pRJM-24 | This study |
| EcRJM-110 | TW14359 ΔgrvA ΔgadE::kan/pRJM-26 | This study |
| EcRJM-83 | TW14359 ΔgrvA/pRJM-22 | This study |
| EcRJM-84 | TW14359 ΔrcsB ΔgrvA/pRJM-22 | This study |
| EcRJM-85 | TW14359 ΔrcsB ΔgrvA/pRJM-20 | This study |
| EcRJM-86 | TW14359 rcsDB-luxE | This study |
| EcRJM-87 | TW14359 grvAB-luxE | This study |
| EcRJM-88 | TW14359 rcsDB-luxE/pluxCDAB3 | This study |
| EcRJM-89 | TW14359 grvAB-luxE/pluxCDAB3 | This study |
| EcRJM-90 | TW14359 ΔgadE::kan | This study |
| EcRJM-91 | TW14359 ΔgrvA ΔgadE::kan | This study |
| EcRJM-92 | TW14359 ΔgadE::kan/pRJM-30 | This study |
| EcRJM-93 | TW14359 ΔgrvA ΔgadE::kan/pRJM-30 | This study |
| EcRJM-94 | TW14359 zinT-FLAG::kan | This study |
| EcRJM-95 | TW14359 ΔgrvA zinT-FLAG::kan | This study |
| EcRJM-96 | TW14359/pRJM-31 | This study |
| EcRJM-97 | EPECa E2348/69/pRJM-31 | This study |
| EcRJM-98 | MG1655/pRJM-31 | This study |
| EcRJM-99 | DH5α/pRJM-31 | This study |
| EcRJM-100 | TW14359/pRJM-32 | This study |
| EcRJM-101 | EPEC E2348/69/pRJM-32 | This study |
| EcRJM-102 | MG1655/pRJM-32 | This study |
| EcRJM-103 | DH5α/pRJM-32 | This study |
| EcRJM-104 | TW14359 rcsDB-luxE/pluxCDAB3/pRJM-21 | This study |
| EcRJM-105 | TW14359 grvAB-luxE/pluxCDAB3/pRJM-21 | This study |
| EcRJM-106 | TW14359 ΔgrvA/pRJM-23 | This study |
| EcRJM-107 | TW14359 ΔgrvA/pRJM-24 | This study |
| EcRJM-108 | TW14359 ΔgrvA/pRJM-25 | This study |
| EcRJM-109 | TW14359 ΔgrvA/pRJM-26 | This study |
| Plasmids | ||
| pACYC177 | Low-copy-no. cloning vector, Ampr Kanr P15A | |
| pBAD22 | Ara-inducible expression vector, Ampr M13 | 92 |
| pMPM-A2 | Low-copy-no. cloning vector, Ampr pMB1/f1 | 36 |
| pMPM-K3 | Low-copy-no. cloning vector, Kanr p15A/f1 | 36 |
| pMPM-T3 | Low-copy-no. cloning vector, Tetr p15A/f1 | 36 |
| pUC19 | High-copy-no. cloning vector, Ampr pMB1 | 93 |
| pRS551 | lac fusion vector, Ampr Kanr lacZ+ ColE1 | |
| pCP20 | FLP recombinase expression vector | 34 |
| pKD4 | Template plasmid for Kan cassette | 34 |
| pKM208 | Bacteriophage λ Red recombinase expression vector | 34 |
| pET-24d | T7 promoter, His tag vector, Kanr f1, pBR322 | Novagen |
| pSU312 | FLAG epitope template, Ampr Kanr, R6K | 94 |
| pRJM-20 | pACYC177-rcsB | 20 |
| pRJM-33 | pACYC177-grvAB | This study |
| pRJM-21 | pMPM-K3-rcsDB | This study |
| pRJM-22 | pMPM-A2-grvA | This study |
| pRJM-23 | pRS551-gadEP(−773, −1)b | This study |
| pRJM-24 | pRS551-gadEP(−320, −1) | This study |
| pRJM-25 | pRS551-gadEP(−516, −276) | This study |
| pRJM-26 | pRS551-gadEP(−773, −497) | This study |
| pRJM-27 | pET-24d-6×His-rcsB | This study |
| pRJM-28 | pMPM-T3-kan | This study |
| pRJM-29 | pMPM-T3-luxE-kan | This study |
| pRJM-30 | pMPM-A2-gadE | This study |
| pRJM-31 | pRS551-grvAP(−268, +38) | This study |
| pRJM-32 | pUC-grvA-6×His | This study |
| placlux8 | placlux8 | 40 |
| pLuxCDAB3 | pLuxCDAB3 | 40 |
a
EPEC, enteropathogenic E. coli.
b
The numbers indicate the positions in gadEP.