Table 3 Bacterial strains, plasmids and oligonucleotides used in this study
Strain, plasmid, oligonucleotide | Relevant genotype/characteristics/information or sequencea | Source/reference |
|---|---|---|
E. coli | Â | Â |
DH5αMCR | F- endA1 supE44 mcrA thi-1 λ-recA1 gyrA96 relA1 deoR Δ(lacZYA-argF) U169 (Φ80dlacZ ΔM15) Δ(mrr-hsdRMS-mcrBC) | [104] |
C. glutamicum | Â | Â |
ATCC 13032 | Nxr, wild-type | ATCC b |
RES167 | Nxr, Δ(cglIMR, cglIIR) Restriction deficient mutant of C. glutamicum ATCC 13032 | [79] |
CG281 | Δcg0910 | This study |
Transposon vector | Â | Â |
pAT6100 | RP4mob, oriVE.c., Kmr Mobilizable cloning vector pK18mob2 carrying the IS6100 insertion element | [31] |
Plasmids | Â | Â |
pK18mobsacB | sacB, lacZα, mcs, Kmr/mobilizable E. coli cloning vector, allows selection for double-crossover in C. glutamicum | [5] |
pK18mobsacB-Δcg0910 | pK18mobsacB with defined deletion derivative of cg0910 | This study |
pEC-XT99A | Tcr, lacIq, mcs, P trc inducible E. coli – C. glutamicum shuttle expression vector | [52] |
pEC-XT99A-cg0910 ex | pEC-XT99A with internal cg0910 exc fragment | This study |
Oligonucleotides 5'-3' | Â | Â |
IS6100x | GGTACAGGTAGGTCCACTTG | This study |
IS6100y | CGGCAGGTGAAGTATCTCAA | This study |
gs_trpP | CACCTGCATCAAGGTCGATT | This study |
s6100e | GCGCCTTGTGGAGAGAGCTT | This study |
s6100x | CGGATAGCGACAATACCAGC | This study |
cg0910_del1 | GATCTAGGATCCGATGGCACCTACAACTTCAC | This study |
cg0910_del2 | AACGATCCAGCGTCTCATCGTCGGCAAGTTCGGCAAGTTC | This study |
cg0910_del3 | CGATGAGACGCTGGATCGTT | This study |
cg0910_del4 | GATCTACTCGAGAGCTCTTCCAGCGTTCCATT | This study |
cg0910_ex1 | GATCTACAATTGAAAGGAGGACAACCATGAGCAAATATGCAGACGA | This study |
cg0910_ex2 | GATCTAGGATCCCTATTTTAAACGATCCAGCG | This study |