TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | Molecular cloning strain | BRL, Gaithersburg, MD |
| SM10λpir | Conjugation strain | 43 |
| B. bronchiseptica | ||
| RB50 | Wild-type B. bronchiseptica | 9 |
| SP5 | RB50 containing an in-frame deletion of codons 227-756 of prn | 13 |
| RBXQ7 | RB50 in which codon 265 of prn was changed to encode glutamic acid instead of aspartic acid (PRN D265E) | This study |
| RB545 | RB50 containing deletions of codons 269-288 and 578-606 in prn (PRNΔR1ΔR2) | This study |
| SP5::pVC4 | SP5 with plasmid pVC4 integrated into the chromosome as shown in Fig. 1A | This study |
| RB50::PRN-NT-HA | RB50 with plasmid pPRN-NT-HA integrated into the chromosome | This study |
| RB545::PRN-NT-HA | RB545 with plasmid pPRN-NT-HA integrated into the chromosome | This study |
| RBXQ7::PRN-NT-HA | RBXQ7 with plasmid pPRN-NT-HA integrated into the chromosome | This study |
| RBX9 | RB50 containing an in-frame deletion of codons 5-3647 in fhaB | 11 |
| RBX26 | RB50 containing an in-frame deletion of codons 227-756 of prn and an in-frame deletion of codons 5-3647 in fhaB | This study |
| Plasmids | ||
| pEG7 | pBR322-based suicide plasmid (Apr and Gmr) | 10 |
| pSS4245 | pBR322-based allelic exchange plasmid (see Fig. 1 and text) | This study |
| pVC1 | pSS4245 containing a 1-kb DNA fragment from prn with codon 265 changed to encode glutamic acid instead of aspartic acid | This study |
| pCI78a | pSS4245 containing a 1-kb DNA fragment from prn with codons 578-606 deleted from the center | This study |
| pXQ4 | pSS4245 containing a 1-kb DNA fragment from prn with codons 269-288 deleted from the center | This study |
| pVC4 | pEG7 containing the entire prn gene and its promoter region | This study |
| pPRN-NT-HA | pEG7 containing a 0.6-kb fragment corresponding to the 5′ end of prn with nucleotides encoding an HA epitope inserted between codons 38 and 39 (corresponding to the first and second amino acids of the mature PRN protein following cleavage of the signal peptide) | This study |