Table 1.
Bacterial strains, plasmids, and primers in this study
| Strain, plasmid, or primer | Characteristic(s)a or sequence (5′–3′) | Reference, source, and/or function |
|---|---|---|
| Strains | ||
| Paenibacillus sp. strain NyZ101 | Wild type; 3-hydoxybenzoate+ gentisate+ | This study |
| E. coli DH5α | supE44 lacU169 (ϕ80lacZΔM15) recA1 endA1 hsdR17 Δthi gyrA96 relA1 | 42 |
| E. coli BL21(DE3) | F− ompT hsdS (rB− mB−) gal dcm lacY1(DE3) | Novagen |
| Plasmids | ||
| pGEM-T | Apr; cloning vector | Promega |
| pET-28a | Kmr; expression vector | Novagen |
| pZWT50 | NdeI-BamHI fragment containing bagI inserted into pET28a | This study |
| pZWT51 | NdeI-EcoRI fragment containing bagL inserted into pET28a | This study |
| pZWT52 | NdeI-EcoRI fragment containing bagK inserted into pET28a | This study |
| pZWT53 | SacI-XhoI fragment containing bagX inserted into pET28a | This study |
| pZWTH46 | pZWT51, in which CAT code for His46 was changed to GCT (alanine) | This study |
| pZWTH143 | pZWT51, in which CAT code for His143 was changed to GCT (alanine) | This study |
| pZWTH147 | pZWT51, in which CAT code for His147 was changed to GCT (alanine) | This study |
| Primers | ||
| g1 | GACTGGATCCATATGGCAGAAAAAATGGAAT | To amplify the bagI gene |
| g2 | ACATGGATCCTAGAATAGGATGGATGAAAG | |
| m1 | GGAGAATTCATATGAGTATTCAGGAAGCGA | To amplify the bagL gene |
| m2 | AGGTGAATTCTGGTACTTCTCGATTTCCT | |
| f1 | CGACGAATTCATATGAAACTAGTGAGCTTTCG | To amplify the bagK gene |
| f2 | TAGAGAATTCGGATCGCTTCCTGAATACTCA | |
| h1 | ACTGAGCTCGCATTGCTTTCATCCATCCT | To amplify the bagX gene |
| h2 | CTGCTCGAGATGTTTTCGTATAGCCGGTT | |
| RTFM | GGCGACATAGTCCGTATTGAA | To amplify 219 bp of the bagK-bagL spanning region |
| CATGGCACAGCACCTCGATAA | ||
| RTMG | CCATCTGCTGGTCGAGCATCT | To amplify 219 bp of the bagK-bagL spanning region |
| CAGGGATGGCATTCCAGAGAG | ||
| RTGH | TCTTCTCTGTCAGCGATCTG | To amplify 207 bp of bagL-bagI spanning region |
| CCAGCACTTGAACGGATTTG | ||
| RT16S | GCTTGAGTGCAGGAGAGGAAA | To amplify a 110-bp fragment of 16S rDNA |
| TCAGCGTCAGTTACAGTCCAG |
a
+, The ability to grow on the indicated substrates. Apr, ampicillin resistance; Kmr, kanamycin resistance.