TABLE 1.
Plasmids and bacterial strains
| Plasmid or strain | Relevant featurea | Source or reference |
|---|---|---|
| Plasmids | ||
| pIH120 | pUC18 carrying the entire F18 fimbria-encoding region | 20 |
| pMAL-p2 | Apr; E. coli expression vector | 12; NEB |
| pKTH5068 | FedFΔ1-10-encoding region in pMAL-p2 | This study |
| pKTH5069 | FedFΔ1-37-encoding region in pMAL-p2 | This study |
| pKTH5070 | FedFΔ1-52-encoding region in pMAL-p2 | This study |
| pKTH5071 | FedFΔ70-77-encoding region in pMAL-p2 | This study |
| pKTH5072 | FedFΔ88-97-encoding region in pMAL-p2 | This study |
| pKTH5073 | FedFΔ110-117-encoding region in pMAL-p2 | This study |
| pKTH5075 | FedFΔ124-280-encoding region in pMAL-p2 | This study |
| pKTH5076 | FedFΔ192-280-encoding region in pMAL-p2 | This study |
| pKTH5077 | FedFΔ247-280-encoding region in pMAL-p2 | This study |
| pKTH5094 | FedF60-123-encoding region in pMAL-p2 | This study |
| pKTH5104 | FedF (Lys72→Ala) mutation in pIH120 | This study |
| pKTH5105 | FedF (Arg75→Ala) mutation in pIH120 | This study |
| pKTH5106 | FedF (Glu76→Ala) mutation in pIH120 | This study |
| pKTH5107 | FedF (Glu82→Ala) mutation in pIH120 | This study |
| pKTH5108 | FedF (Arg84→Ala) mutation in pIH120 | This study |
| pKTH5109 | FedF (His88→Ala) mutation in pIH120 | This study |
| pKTH5110 | FedF (His89→Ala) mutation in pIH120 | This study |
| pKTH5111 | FedF (Glu96→Ala) mutation in pIH120 | This study |
| pKTH5124 | pIH120/FedFΔ1-5 | This study |
| pKTH5125 | pIH120/FedFΔ1-34 | This study |
| pKTH5126 | pIH120/FedFΔ1-52 | This study |
| pKTH5127 | pIH120/FedFΔ70-77 | This study |
| pKTH5128 | pIH120/FedFΔ88-97 | This study |
| pKTH5129 | pIH120/FedFΔ110-117 | This study |
| pKTH5130 | pIH120/FedFΔ124-280 | This study |
| pKTH5131 | pIH120/FedFΔ192-280 | This study |
| pKTH5132 | pIH120/FedFΔ247-280 | This study |
| pKTH5149 | FedF (Lys72, His88, His89→Ala) triple mutation in pIH120 | This study |
| pKTH5150 | FedF (Lys72, His88→Ala) double mutation in pIH120 | This study |
| pKTH5151 | FedF (Lys72, His89→Ala) double mutation in pIH120 | This study |
| pKTH5152 | FedF (His88, His89→Ala) double mutation in pIH120 | This study |
| E. coli strains | ||
| F107/86 | Clinical isolate with F18 fimbriae | 19 |
| DH5αF′ | F′ endA1 hsd17 (rk− mk+) supE44 thi-1 recA1 gyrA (Nalr) relA1Δ(laclZYA-argF) U169 deoR [φ80d lacΔ(lacZ) M15] | 49 |
| HB101 | Hsr− Hsm− RecA− Gal− Pro−str::r F− | 5 |
| ERF2055 | HB101 with pIH120 | 40 |
| ERF2079 | HB101 with pKTH5124 | This study |
| ERF2080 | HB101 with pKTH5125 | This study |
| ERF2081 | HB101 with pKTH5126 | This study |
| ERF2082 | HB101 with pKTH5127 | This study |
| ERF2083 | HB101 with pKTH5128 | This study |
| ERF2084 | HB101 with pKTH5129 | This study |
| ERF2085 | HB101 with pKTH5130 | This study |
| ERF2086 | HB101 with pKTH5131 | This study |
| ERF2087 | HB101 with pKTH5132 | This study |
| ERF2088 | DH5αF′ with pKTH5068 | This study |
| ERF2089 | DH5αF′ with pKTH5069 | This study |
| ERF2090 | DH5αF′ with pKTH5070 | This study |
| ERF2091 | DH5αF′ with pKTH5071 | This study |
| ERF2092 | DH5αF′ with pKTH5072 | This study |
| ERF2093 | DH5αF′ with pKTH5073 | This study |
| ERF2094 | DH5αF′ with pKTH5075 | This study |
| ERF2095 | DH5αF′ with pKTH5076 | This study |
| ERF2096 | DH5αF′ with pKTH5077 | This study |
| ERF2107 | HB101 with pUC18 | This study |
| ERF2114 | DH5αF′ with pKTH5094 | This study |
| ERF2121 | HB101 with pKTH5104 | This study |
| ERF2122 | HB101 with pKTH5105 | This study |
| ERF2123 | HB101 with pKTH5106 | This study |
| ERF2124 | HB101 with pKTH5107 | This study |
| ERF2125 | HB101 with pKTH5108 | This study |
| ERF2126 | HB101 with pKTH5109 | This study |
| ERF2127 | HB101 with pKTH5110 | This study |
| ERF2128 | HB101 with pKTH5111 | This study |
| ERF2143 | HB101 with pKTH5149 | This study |
| ERF2144 | HB101 with pKTH5150 | This study |
| ERF2145 | HB101 with pKTH5151 | This study |
| ERF2146 | HB101 with pKTH5152 | This study |
a
Numbers following “FedFΔ” represent positions of amino acids deleted from the mature FedF protein.