TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Descriptiona | Referenceb |
|---|---|---|
| B. methanolicus strains | ||
| MGA3 | Wild-type strain, MeOH+ | 33 |
| NOA2 | Wild-type strain, MeOH+ | 33 |
| PB1 NCIMB 13113 | Wild-type strain, MeOH+ | NCIMB |
| HEN9 | Wild-type strain, MeOH+ | R. Dillingham |
| TSL32 | Wild-type strain, MeOH+ | R. Dillingham |
| DFS2 | Wild-type strain, MeOH+ | R. Dillingham |
| CFS | Wild-type strain, MeOH+ | R. Dillingham |
| RCP | Wild-type strain, MeOH+ | R. Dillingham |
| SC6 | Wild-type strain, MeOH+ | R. Dillingham |
| NIWA | Wild-type strain, MeOH+ | R. Dillingham |
| BVD | Wild-type strain, MeOH+ | R. Dillingham |
| DGS | Wild-type strain, MeOH+ | R. Dillingham |
| JCP | Wild-type strain, MeOH+ | R. Dillingham |
| N2 | Wild-type strain, MeOH+ | R. Dillingham |
| MGA3C-A6 | MGA3 cured of pBM19, MeOH− | This study |
| E. coli strain | ||
| DH5α | General cloning host | BRL |
| Plasmids | ||
| pUC19 | General cloning vector, Apr | NEB |
| pGEM-3zf | General cloning vector, Apr | Promega |
| pGEM-11zf | General cloning vector, Apr | Promega |
| pLITMUS28 | General cloning vector, Apr | NEB |
| pDQ508 | E. coli-B. methanolicus shuttle plasmid, Neor Apr | 13 |
| pJB658 | Low-copy-number cloning vector, Apr | 9 |
| pBM19 | Endogenous B. methanolicus plasmid | This study |
| pTB1.9 | E. coli-B. methanolicus shuttle vector, Neor Apr | This study |
| pTB3.3H | 3,289-bp HindIII fragment from pBM19 cloned into pGEM-3zf | This study |
| pTB1.9mdh | pTB1.9 harboring mdh gene and 237 bp of upstream sequence | This study |
| pTB1.9mdhL | pTB1.9 harboring mdh gene and 1,125 bp of upstream sequencec | This study |
| pRMP1 | 910-bp PCR fragment encompassing the 3′ end of hps and the 5′ end of phi cloned into the XbaI/NcoI sites of pLITMUS28 | This study |
| pRMP2 | 1,056-bp PCR fragment encompassing the hps gene and upstream regions cloned into the HindIII/SalI sites of pGEM-11zf | This study |
| pRMP3 | 920-bp PCR fragment encompassing the 3′ end of hps and the phi gene cloned into the HindIII/SalI sites of pGEM-11zf | This study |
a
Apr, ampicillin resistance; Neor, neomycin resistance; MeOH+, grows on methanol; MeOH−, does not grow on methanol.
b
NCIMB, National Collection of Industrial and Marine Bacteria; BRL, Bethesda Research Laboratories; NEB, New England Biolabs.
c
See Materials and Methods.