Table 2.
List of bacterial strains and plasmids used in this study
| Strain, plasmid | Function and relevant characteristics | References |
|---|---|---|
| B. methanolicusstrains | ||
| MGA3 | Wild type strain, ATCC 53907 | [53] |
| PB1 | Wild type strain, NCIMBI 13113 | [53] |
| E. colistrains | ||
| DH5α | General cloning host. F−thi-1 endA1 hsdR17(r− m−) supE44 ΔlacU169 (−80lacZΔM15) recA1 gyrA96 relA1 | Bethesda Research Laboratories |
| BL21 | protein expression host. ompT hsdSB (rB− mB_) gal dcm (DE3) | Novagen |
| C. glutamicumstrains | ||
| ATCC 13032 | WT strain, auxotrophic for biotin | [1] |
| Δtal | In-frame deletion of the tal gene of WT | This study |
| Plasmids | ||
| pVWEx1 | KmR; C. glutamicum-E. coli shuttle vector (PtaclacIqoriVC.goriVE.c) | [46] |
| pVWEx1-taPB1 | derived from pVWEx1, for regulated expression of tal of B. methanolicus PB1 | This study |
| pVWEx1-taMGA3rec | derived from pVWEx1, for regulated expression of modified tal of B. methanolicus MGA3 | This study |
| pVWEx1-talCG | derived from pVWEx1, for regulated expression of tal (cg1776) of C. glutamicum ATCC 13032 | This study |
| pHP13 | B. subtilis-E coli shuttle vector; ClmR | [22] |
| pTH1mp-taPB1 | pHP13 derivate with tal of B. methanolicus PB1 under control of mdh promoter | This study |
| pET28b | KanR; T7lac; vector for his-tagged protein overproduction | (Novagen) |
| pET28b-taPB1 | purification of his-tagged B. methanolicus PB1 Ta from E. coli BL21(DE3) | This study |
| pET28b-taMGA3rec | purification of his-tagged modified B. methanolicus MGA3 Ta from E. coli BL21(DE3) | This study |
| pET28b-taMGA3_M1 | derived from pET28b, for expression of variant M1 of taMGA3 from B. methanolicus MGA3, see Table 3 | This study |
| pET28b-taMGA3_M2 | derived from pET28b, for expression of variant M2 of taMGA3 from B. methanolicus MGA3, see Table 3 | This study |
| pET28b-taMGA3_M3 | derived from pET28b, for expression of variant M3 of taMGA3 from B. methanolicus MGA3, see Table 3 | This study |
| pET28b-taMGA3_M4 | derived from pET28b, for expression of variant M4 of taMGA3 from B. methanolicus MGA3, see Table 3 | This study |
| pET28b-taMGA3_M5 | derived from pET28b, for expression of variant M5 of taMGA3 from B. methanolicus MGA3, see Table 3 | This study |
| pET28b-taMGA3_M6 | derived from pET28b, for expression of variant M6 of taMGA3 from B. methanolicus MGA3, see Table 3 | This study |
| pET28b-taMGA3 | derived from pET28b, for expression of variant of the annotated taMGA3 from B. methanolicus MGA3, see Table 3 | |
| pK19mobsacB | KanR; vector for gene deletions (RP4 mob; sacB B. subtilis; lacZ; OriVE.c.) | [52] |
| pK19mobsacB-Δtal | derived from pK19mobsacB for in-frame deletion of C. glutamicum tal | This study |
Abbreviations: SpeR, spectinomycin resistance; CmR, chloramphenicol resistance; KanR kanamycin resistance