TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant genotype or description | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | φ80dlacZΔM15 ΔlacU169 deoR recA endA1 | Life Technologies |
| BL21(DE3) | hsdS gal (λ cI ts857 ind1 Sam7 nin5 lacUV5-T7 gene 1) | 18 |
| H. influenzae strains | ||
| C54b−p−Hsf+ | Capsule-deficient, nonpiliated mutant of type b strain C54 | 5 |
| C54b−p−Hsf− | Derivative of C54b−p−Hsf+ containing a mini-Tn10 kan element in the hsf locus | 5 |
| Plasmids | ||
| pT7-7 | Cloning vector, Ampr | New England Biolabs |
| pDC601 | pT7-7 containing hsf from H. influenzae strain C54 | 15 |
| pDC601(E569A) | pDC601 encoding E-to-A mutation at residue 569 in Hsf | 3 |
| pDC601(D1935A) | pDC601 encoding D-to-A mutation at residue 1935 in Hsf | 3 |
| pDC601(E569A/D1935A) | pDC601 encoding E-to-A and D-to-A mutations at residues 569 and 1935 in Hsf | 3 |
| pNS1 | Derivative of pHAT10 that contains hia upstream sequence and coding sequence for the Hia signal peptide, the HAT epitope, and coding sequence for the Hia translocator domain | 3 |
| pNS1-HsfBD1 | pNS1 containing coding sequence for HsfBD1 from H. influenzae strain C54 | This study |
| pNS1-HsfBD2 | pNS1 containing coding sequence for HsfBD2 from H. influenzae strain C54 | This study |
| pNS1-HsfBD1(Y1934V) | pNS1/HsfBD1 encoding Y-to-V mutations at residue 1934 in Hsf | This study |
| pNS1-HsfBD1(D1935E) | pNS1/HsfBD1 encoding D-to-E mutation at residue 1935 in Hsf | This study |
| pNS1-HsfBD2(V568Y) | pNS1/HsfBD2 encoding V-to-Y mutation at residue 568 in Hsf | This study |
| pNS1-HsfBD2(E569D) | pNS1/HsfBD2 encoding E-to-D mutation at residue 569 in Hsf | This study |
| pGEX-6-P-1/HsfBD1 | pGEX-6-P-1 containing coding sequence for HsfBD1 from H. influenzae strain C54 | 3 |
| pGEX-6-P-1/HsfBD2 | pGEX-6-P-1 containing coding sequence for HsfBD2 from H. influenzae strain C54 | 3 |
| pGEX-6-P-1/HsfBD1(Y1934) | Derivative of pGEX-6-P-1/HsfBD1 encoding Y-to-V mutation at residue 1934 in Hsf | This study |
| pGEX-6-P-1/HsfBD1(D1935E) | Derivative of pGEX-6-P-1/HsfBD1 encoding D-to-E mutation at residue 1935 in Hsf | This study |
| pGEX-6-P-1/HsfBD2(V568Y) | Derivative of pGEX-6-P-1/HsfBD2 encoding V-to-Y mutation at residue 568 in Hsf | This study |
| pGEX-6-P-1/HsfBD2(E569D) | Derivative of pGEX-6-P-1/HsfBD2 encoding E-to-D mutation at residue 569 in Hsf | This study |