Skip to main content
. 2008 Dec 1;75(2):419–427. doi: 10.1128/AEM.01844-08

TABLE 1.

Strains, plasmids, and oligonucleotides used in this study

Strain, plasmid, or oligonucleotide Relevant characteristics or sequence Source, reference, or purpose
Strains
    E. coli DH5α supE44 hsdR17 recA1 endA1 gyrA96 thi-1 relA1 17
    WT C. glutamicum WT strain ATCC 13032, biotin-auxotrophic American Type Culture Collection
    C. glutamicum DM1729 pyc(P458S) hom(V59A) lysC(T311I), derived from WT C. glutamicum For construction, see Table S1 in the supplemental material
    C. glutamicum DM1729 ΔilvB Strain DM1729 with deletion of ilvB, encoding the large subunit of the acetolactate synthase (AHAS) This work
    C. glutamicum DM1729 ΔC-T ilvN Strain DM1729 with deletion of the last 83 amino acids of the C-terminal domain of ilvN, encoding the small subunit of the AHAS This work
    C. glutamicum DM1933 Δpck pyc(P458S) hom(V59A), 2 copies of lysC(T311I), 2 copies of asd, 2 copies of dapA, 2 copies of dapB, 2 copies of ddh, 2 copies of lysA, 2 copies of lysE derived from WT C. glutamicum For construction, see Table S1 in the supplemental material
    C. glutamicum DM1933 ΔilvB Strain DM1933 with deletion of ilvB, encoding the large subunit of the AHAS This work
    C. glutamicum DM1933 ΔC-T ilvN Strain DM1933 with deletion of the last 83 amino acids of the C-terminal domain of ilvN, encoding the small subunit of the AHAS This work
Plasmids
    pK19mobsacB Kmr, mobilizable (oriT), oriV 44
    pK19mobsacB ΔilvB pK19mobsacB carrying a truncated ilvB gene 32
    pK19mobsacB ΔC-T ilvN pK18mobsacB carrying a truncated ilvN gene (shortened by 249 bp) This work
Oligonucleotides
    P1 5′-CCCAAGCTTGCTGTTTCCAGATGACCAACC-3′ Primer for deletion of ΔC-T ilvN
    P2 5′-GGCGATAGTGGTCTCTTCATCAAGTCGCACGACTTTGAGC-3′ Primer for deletion of ΔC-T ilvN; crossover overlap underlined
    P3 5′-GAAGAGACCACTATCGCCACAGCAATTAATCTGATTGC-3′ Primer for deletion of ΔC-T ilvN; crossover overlap underlined
    P4 5′-CGCGGATCCCGTTCAGGTTTGGCTCGATG-3′ Primer for deletion of ΔC-T ilvN and to verify ilvB deletion
    PD1 5′-CCAAGATGGCTAATTCTGACGTCACC-3′ Primer to verify ΔC-T ilvN deletion
    PD2 5′-GACTAGTCACATTTATGCAGCAGGTGC-3′ Primer to verify ΔC-T ilvN deletion
    PilvB 5′-GCAACAGACATCTGTCGC-3′ Primer to verify ilvB deletion