Table 1.
Plasmids and bacterial strains used in these studies
| Strain or plasmid | Descriptiona | Reference |
|---|---|---|
| Strains | ||
| H10407 | ETEC (origin, Bangladesh) O78:H11 CFA/I LT/ST-1a/ST-1b | 61 |
| E24377A | ETEC (origin, Egypt) O139:H28 CFA/II (CS1/CS3) LT/ST | 62 |
| BL21-AI | F− ompT hsdSB(rB− mB−) gal dcm araB::T7 RNAP-tetA | |
| GPM1746a | H10407 hsdR1::lacZ::Tn10 | G. P. Munson, unpublished data |
| jf2624 | GPM1746a containing single-copy integrant of eaeH promoter-lacZYA transcriptional fusion at attBHK022 | This study |
| jf2575 | GPM1746a containing promoterless lacZYA reporter integrated at attBHK022 | This study |
| jf2450 | H10407(pGFPmut3.1) | This study |
| Plasmids | ||
| pAH69 | CRIM integrase expression helper plasmid | 23 |
| pHKLac1 | Reporter with promoterless lacZYA, R6Kγ, aadA, attPHK022 | 22 |
| pQL002 | eaeH promoter cloned into BamHI/EcoRI sites of pHKLac1 | This study |
| pGFPmut3.1 | pUC18-based gfpmut3* expression plasmid, Ampr | 63 |
| pDONOR221 | Entry cloning plasmid containing ccdB negative-selection gene and Cmr cassette between lambda attP sites for recombination with PCR products containing attB regions, Kmr | Invitrogen |
| pET-DEST42 | T7 lac promoter/IPTG-inducible expression plasmid, ccdB, Cmr genes flanked by attR sites for recombination with entry plasmid, C-terminal V5 and 6-His tags | Invitrogen |
| pSS001 | 4,254-bp eaeH amplicon cloned into pDONOR221 | This study |
| pSS002 | EaeH expression plasmid derived from LR recombination reaction of pSS001 and pET-DEST42 placing eaeH in frame with V5 and 6-His epitope tags | This study |
a
Ampr, ampicillin resistance; Kmr, kanamycin resistance; Cmr, chloramphenicol resistance; aadA, aminoglycoside-3-adenyltransferase gene encoding streptomycin and spectinomycin resistance; attB, bacterial chromosome phage integration site; attP, complementary phage-derived integration site; R6Kγ, pir-dependent origin of replication.