TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial strain | Description | Source or reference |
|---|---|---|
| M. catarrhalis | ||
| O35E | Wild-type isolate | 2 |
| ETSU-9 | Wild-type isolate | Steven Berk |
| ETSU-9.Strr | Spontaneous streptomycin-resistant rpsL mutant of ETSU-9 | 42 |
| ETSU-9.1 | uspA1 null mutant of ETSU-9; kanamycin resistant | This study |
| ETSU-9.1Strr | Streptomycin-resistant ETSU-9.1 | This study |
| ETSU-9.2 | uspA2H null mutant of ETSU-9 constructed using pELU244SPEC as the source of the mutant allele; spectinomycin resistant | This study |
| ETSU-9.12 | uspA1 uspA2H mutant of ETSU-9; kanamycin and spectinomycin resistant | This study |
| ETSU-9.12Strr | Streptomycin-resistant ETSU-9.12 | This study |
| ETSU-9.2H | uspA2H null mutant of ETSU-9 constructed using pMPGTG3spec as the source of the mutant allele; spectinomycin resistant | This study |
| ETSU-9.mut92 | ETSU-9 mutant with 418-aa deletion in the UspA2H protein | This study |
| ETSU-9.mut92.2 | uspA2H null mutant of ETSU-9mut.92 | This study |
| ETSU-9.698 | ETSU-9 with a transposon insertion in the uspA2H gene; Kanr | This study |
| ETSU-9.724 | ETSU-9 with a transposon insertion in the uspA2H gene; Kanr | This study |
| ETSU-9.2311 | ETSU-9 with a transposon insertion in the uspA2H gene; Kanr | This study |
| ETSU-9.222 | ETSU-9 with a transposon insertion in the putative lytic murein transglycosylase D gene; Kanr | This study |
| ETSU-9.222cat | Lytic murein transglycosylase D gene mutant of ETSU-9; Cmr | This study |
| ETSU-9.714 | ETSU-9 with a transposon insertion in the putative aminoglycoside phosphotransferase gene; Kanr | This study |
| ETSU-9.714cat | Aminoglycoside phosphotransferase mutant of ETSU-9; Cmr | This study |
| ETSU-9.988 | ETSU-9 with a transposon insertion in the putative ampD gene; Kanr | This study |
| ETSU-9.988cat | ampD mutant of ETSU-9; Cmr | This study |
| E. coli | ||
| EPI3000 | Host for CopyControl pCC1 vector | Epicentre |
| DH5α | Host for cloning experiments | Invitrogen |
| E. coli INVαF′ | Host for cloning experiments | Invitrogen |
| Plasmids | ||
| pCR-Blunt | Cloning vector; Kanr Zeocinr | Invitrogen |
| pMPGTG3 | pCR-Blunt containing the majority of the ETSU-9 uspA2H ORF | This study |
| pMPGTG3spec | pMPGTG3 with a spectinomycin resistance cartridge inserted in the uspA2H ORF | This study |
| pELU244SPEC | pBluescript containing a truncated uspA2 gene from M. catarrhalis P44 into which a spectinomycin resistance cartridge was inserted | 22 |
| pCR2.1 | Cloning vector; Kanr Ampr | Invitrogen |
| pMP222 | pCR2.1 containing the putative M. catarrhalis lytic murein transglycosylase D gene | This study |
| pMP222cat | pMP222 with a promoterless cat cartridge inserted into the NdeI site in the putative lytic murein transglycosylase D gene; Kanr Ampr Cmr | This study |
| pMP714 | pCR2.1 containing the putative M. catarrhalis aminoglycoside phosphotransferase gene | This study |
| pMP714cat | pMP714 with a promoterless cat cartridge inserted into the AccI site in the putative aminoglycoside phosphotransferase gene; Kanr Ampr Cmr | This study |
| pMP988 | pCR2.1 containing the putative M. catarrhalis ampD gene | This study |
| pMP988cat | pMP988 with a promoterless cat cartridge inserted into the AccI site in the ampD gene; Kanr Ampr Cmr | This study |
| pUSPA1KAN | pBluescript with a uspA1 gene fragment containing a kan cartridge | 2 |
| pCC1 | CopyControl vector; Cmr | Epicentre |
| pCC1kan | pCC1 containing a cloned kan cartridge | This study |
| pMPE9U2H | pCC1 containing the wild-type ETSU-9 uspA2H gene | This study |