Table 1.
List of strains and plasmids used
| Strain, plasmid | Function and relevant characteristics | References |
|---|---|---|
|
B. methanolicus
|
|
|
| MGA3 |
Wild-type strain |
[19] |
|
E. coli
|
|
|
| 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 |
|
|
| pEKEx3 |
SpeR; C. glutamicum/E. coli shuttle vector (P
tac
, lacIq; 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 PciI 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 |
Abbreviations: SpeR, spectinomycin resistance; ClmR, chloramphenicol resistance; AmpR, ampicillin resistance.