Table 3.
Bacterial strains, plasmids and oligonucleotides used in this study
| Strain, plasmid, oligonucleotide | Relevant genotype/characteristics/information or sequence a | Source/reference |
| E. coli | ||
| DH5αMCR | F- endA1 supE44 mcrA thi-1 λ-recA1 gyrA96 relA1 deoR Δ(lacZYA-argF)U169 (Φ80dlacZΔM15) Δ(mrr-hsdRMS-mcrBC) | [104] |
| C. glutamicum | ||
| 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 | ||
| pAT6100 | RP4mob, oriVE.c., Kmr Mobilizable cloning vector pK18mob2 carrying the IS6100 insertion element |
[31] |
| Plasmids | ||
| 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, Ptrc inducible E. coli – C. glutamicum shuttle expression vector |
[52] |
| pEC-XT99A-cg0910ex | pEC-XT99A with internal cg0910 exc fragment | This study |
| Oligonucleotides 5'-3' | ||
| 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 |
a r superscript indicates resistance. Nx, nalidixic acid; Km, kanamycin; Tc, tetracycline.
b ATCC; American Type Culture Collection, Rockville, MD, USA.
c the postfix ex indicates genes preceded by an artificial RBS