TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Reference(s) or source |
|---|---|---|
| B. methanolicus MGA3 | Wild type | 34 |
| E. coli strains | ||
| DH5α | General cloning host | Bethesda Research Laboratories |
| ER2566 | Expression host, carries chromosomal gene for T7 RNA polymerase | New England Biolabs |
| Plasmids | ||
| pTB1.9mdhL | E. coli-B. methanolicus shuttle vector pTB1.9 carrying the mdh gene, Ampr Neor | 4 |
| pHP13 | E. coli-B. methanolicus shuttle vector, Clmr | 15, 17 |
| pHP13mp-dapG | pHP13 carrying the dapG coding region under control of the mdh promoter | This study |
| pHP13mp-lysC | pHP13 carrying the lysC coding region under control of the mdh promoter | This study |
| pHP13mp-yclM | pHP13 carrying the yclM coding region under control of the mdh promoter | This study |
| pET21a | E. coli expression vector, six-His tag, T7 promoter, Ampr | Novagen |
| pSB1 | pET21a with dapG coding sequence under control of the T7 promoter and fused to six-His tag | This study |
| pSB3 | pET21a with lysC coding sequence under control of the T7 promoter and fused to six-His tag | This study |
| pSB4 | pET21a with yclM coding sequence under control of the T7 promoter and fused to six-His tag | This study |
a
Ampr, ampicillin resistance; Neor, neomycin resistance; Clmr, chloramphenicol resistance.