Table 1 List of strains and plasmids used
Strain, plasmid | Function and relevant characteristics | References |
|---|---|---|
B. methanolicus | Â | Â |
MGA3 | Wild-type strain | [19] |
E. coli | Â | Â |
DH5α | F- thi-1 endA1 hsdR17(r - m-) supE44 ΔlacU169 (-80lacZΔM15) recA1 gyrA96 relA1 | Bethesda research labs |
BL21 | ompT hsdSB(rB - mB_) gal dcm (DE3) | Novagen [40] |
Plasmids | Â | Â |
pEKEx3 | SpeR; C. glutamicum/E. coli shuttle vector (P tac , lacI q; pBL1, OriV C.g. , OriV E.c. ) | [41] |
pHP13 | B. methanolicus-E. coli shuttle vector; ClmR | [42] |
pHP13mp | pHP13 carrying lysC coding region under control of the mdh promoter | [39] |
pTH1mp-lysC | Similar as pHP13mp-lysC but with Pci I site upstream mdh promoter removed | [43] |
pTH1mp | pTH1, but with a mdh promoter upstream to the mcs | This work |
pTH1-tkt c (Bme) | Derived from pTH1, for regulated expression of tkt c of B. methanolicus | This work |
pTH1-tkt p (Bme) | Derived from pTH1, for regulated expression of tkt p of B. methanolicus | This work |
pET16b | AmpR; T7lac; vector for his-tagged protein overproduction | (Novagen) |
pET16b-tkt c (Bme) | For production of his-tagged TKTC from B. methanolicus | This work |
pET16b-tkt P(Bme) | For production of his-tagged TKTP from B. methanolicus | This work |
pET16b-gapB | Purification of his-tagged E. coli E4PDH from E. coli BL21(DE3) | This work |