TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Description | Source or reference |
|---|---|---|
| Strains | ||
| E. coli TOP 10 | Cloning strain | Invitrogen |
| M. catarrhalis | ||
| O35E | Serotype A wild-type strain | 29 |
| 26397 | Serotype B wild-type strain | CCUGa |
| 26404 | Serotype C wild-type strain | CCUG |
| D4 | Strain 26404 with EZ::TN transposon insertion in lgt2 gene | This study |
| 26404lgt2 | lgt2 knockout mutant strain | This study |
| Plasmids | ||
| pCR2.1 | TOPO TA cloning vector | Invitrogen |
| pBluescript II KS(+) | Cloning vector | Stratagene |
| pUC4K | Kanamycin resistance cassette | Amersham |
| pCR-lgt2 | lgt2 cloned into pCR2.1 | This study |
| KS-lgt2 | EcoRI-SalI lgt2 fragment cloned into KS(+) | This study |
| KS-lgt2-Kan | EcoRI-blunted kanamycin resistance cassette inserted into blunted HindIII site of KS-lgt2 | This study |
| Primers | ||
| lgt2-F | 5′-CTCGTCGACTGTGAATTTGGCAAACAAGG-3′b | This study |
| lgt2-R | 5′-CTCGAATTCACGCTTACCAATGCTTCGAT-3′c | This study |
| lgt587 | 5′-GCCGAAAATAATCACGCAAC-3′ | This study |
| lgt1482 | 5′-TGGCATCAAAAATCACATGG-3′ | This study |
| lgt1754 | 5′-CCCAAATAGACATCGTCCTCA-3′ | This study |
| lgt2188 | 5′-CCAGCGATGAGTGATGTGTT-3′ | This study |
| lgtR1071 | 5′-CGGTCAAATTGTGCCACATC-3′ | This study |
| lgtR2002 | 5′-TGCCACAAAAAGAAGAGAACA-3′ | This study |
| lgtR3472 | 5′-AAAATTGCCCAAAGTCATGG-3′ | This study |
| KAN-2 RP-1 | 5′-GCAATGTAACATCAGAGATTTTGAG-3′ | Epicenter |
a
CCUG, Culture Collection of the University of Goteborg, Goteborg, Sweden.
b
SalI site underlined.
c
EcoRI site underlined.