TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Descriptiona | Mutagenic oligonucleotide | Source or reference |
|---|---|---|---|
| P. aeruginosa | |||
| FRD1 | Cystic fibrosis isolate; Alg+ | This laboratory | |
| FRD462 | algG4 S272N Alg+ | 4 | |
| FRD1200 | ΔalgG::aacCI | 25 | |
| Plasmids | |||
| pRK2013 | ColE1-Tra(RK2)+ Kmr | 13 | |
| pALTER-Ex1 | Phagemid, Tcr | Promega | |
| pMF54 | Ptrc expression vector pKK233-2 with oriVSForiT laclq Apr | 14 | |
| pMF55 | pMF54 with 2.2 kb algG NcoI-NcoI fragment | 14 | |
| pSJ209 | pMF54 with 1.7 kb algG4 S272N in XbaI-Xho sites | 25 | |
| pSAD1 | pALTER-Ex1 with 2.2 kb algG NcoI-NcoI fragment | This study | |
| pSAD47 | algG H326A | TACGGCATCGACCCGGCCGACCGTTCGCAC | |
| pSAD53 | algG D324A | ATCGTCTACGGCATCCGCCCCGCACGACCCGT | |
| pSAD56 | algG P325A | GTCTACGGCAATCGACGCGCACGACCGTTCGCAC | |
| pSAD68 | algG Y321F | CGCGACAACATCGTCGGCGGCATCGACCCG | |
| pSAD70 | algG D327A | TACGGCATCGACCCGCACGCCCGTTCGCACCGCCT | |
| pSAD84 | algG R316A | AAGGGCAACACCTACGCCGACAACATCGTCTAC | |
| pSAD85 | algG N361A | GACAGCTTCATCTTCGCCAACCGCAGCTACGAG | |
| pSAD86 | algG D317A | GGCAACACCTACCGCGCCAACATCGTCTACGGC | |
| pSAD88 | algG D336A | CGCCTGATCATCGCCGCCAACACCGTCCACGGG | |
| pSAD90 | algG N367A | AACCGCAGCTACGAGGCCAAGCTTTCCGGGATC | |
| pSAD92 | algG N362A | AGCTTCATCTTCAACGCCCGCAGCTACGAGAAC | |
| pSAD102 | algG V383A | AGCGAGGGCAACCTGGCCGCCTACAACGAGGTC | |
| pSAD105 | algG Y385F | GGCAACCTGGTGGCCTTCAACGAGGTCTATCGC | |
| pSAD107 | algG Y365F | TTCAACAACCGCAGCTTCGAGAACAAGCTTTCC | |
| pSAD110 | algG S364A | ATCTTCAACAACCGCGCCTACGAGAACAAGCTT | |
| pSAD137 | algG E366A | AACCGCAGCTACGCGAACAAGCTTTCC | |
| pSAD149 | algG with deletion of amino acids 37-137 | ||
| pSAD151 | algG with deletion of amino acids 37-161 |
a
Abbreviations for phenotypes: Alg+, alginate overproduction; Apr, ampicillin resistance; Kmr, kanamycin resistance; Tra+, transfer by conjugation. Plasmids pSAD47 to pSAD151 contain the gene denoted in the pMF54 vector.