Table 4 List of bacterial strains and plasmids

E. coli

DH5α

General cloning host (F^- thi-1 endA1 hsdR17(r ^- m^-) supE44 ∆lacU169 (^-80lacZ∆M15) recA1 gyrA96 relA1)

BRL

BL21 (DE3)

Host for recombinant protein production (ompT hsdSB(rB ^- mB^_) gal dcm (DE3))

Novagen

C. glutamicum

ATCC13032

WT strain, auxotrophic for biotin

[51]

Δpgk

In-frame deletion of the pgk gene of WT

This work

DM1933

Δpck pyc(P458S) hom(V59A), 2 x of lysC(T311I), 2x of asd, 2x of dapA, 2 x of dapB, 2 x of ddh, 2 x of lysA, 2 x of lysE

[7]

ORN1

L-ornithine overproducing strain derived from WT, auxotrophic for L-arginine due to argF deletion

[6]

ARG1

ΔargR (pEKEx3-argB _{( A26VM31V)}); in-frame deletion of argR

[6]

Plasmids

pGEM-T

General cloning vector

Promega

pEKEx3

Spec^R; C. glutamicum/E. coli shuttle vector (P _tac , lacI ^q; pBL1, OriV _C.g. , OriV _E.c. )

[57]

pEKEx3-pgk (Cg)

Derived from pEKEx3, for regulated expression of pgk of C. glutamicum

This work

pVWEx1

Kan^R, Ptac, lacI^q

[58]

pVWEx1-pgk

Derived from pVWEx1, for regulated expression of pgk of C. glutamicum

This work

pET16b

Amp^R; T7lac; vector for his-tagged protein overproduction

Novagen

pET16b-pgk (Cg)

Purification of his-tagged (His₆) C. glutamicum PGK from E. coli BL21(DE3)

This work

pK19mobsacB

Km^R; E. coli/C. glutamicum shuttle vector for construction of insertion and deletion mutants in C. glutamicum (pK18 oriV _Ec sacB lacZ α)

[59]

pK19mobsacB∆pgk

pK19mobsacB with a pgk deletion construct

This work