Table 2.
List of bacterial strains, plasmids, and oligonucleotides
| Strain, plasmid, or oligonucleotide | Relevant characteristics or sequence | Source, reference, or purpose |
|---|---|---|
| Strains | ||
| Escherichia coli DH5α | F− Φ80lacZΔM15 Δ(lacZYA-argF) U169 endA1 recA1 hsdR17 (rk−, mk+) supE44 thi-1 gyrA96 relA1 phoA | [32] |
| Corynebacterium glutamicum ATCC 13032 | Wild type | American-type culture collection |
| C. glutamicum Δpqo ΔaceE ΔldhA Δmdh | C. glutamicum carrying genetic deletions of the pyruvate:quinone oxidoreductase (pqo), the E1 subunit of the pyruvate dehydrogenase complex (aceE), the lactate dehydrogenase (ldh), and the malate dehydrogenase (mdh) | [33] |
| PDO1 | C. glutamicum + pJULgldA | This study |
| PDO2 | C. glutamicum Δpqo ΔaceE ΔldhA Δmdh + pJULgldA | This study |
| Plasmids | ||
| pJC4 | [34] | |
| pJULgldA | pJC4::(Ptuf–gldA–TrrnB) plasmid expressing the E. coli glycerol dehydrogenase (gldA) under control of the constitutive C. glutamicum EF-TU promoter (Ptuf) and closed by the E. coli rrnB terminator (TrrnB) | This study |
| Oligonucleotides | 5′ → 3′ | |
| P1 | GACGCCGCAGGG TCTAGACCACAGGGTAGCTGGTAGTTTG | Fw primer Ptuf (pJC4, Xba I) |
| P2 | CATGGTATGTCCTCCTGGACTTC | Rv primer Ptuf |
| gldA1 | GAAGTCCAGGAGGACATACCATGGACCGCATTATTCAATC | Fw primer gldA (P tuf) |
| gldA2 | CTTCTCTCATCCGCCAAAACAGCAGGCAATTTTGCGTTC | Rv primer gldA (T rrnB) |
| T1 | CTGTTTTGGCGGATGAGAGAAG | Fw primer TrrnB |
| T2 | GATATCCATCACACTG GCGGCCGCAGGAGAGCGTTCACCGACAAAC | Rv primer TrrnB (pJC4, Not I) |
| seq 1 | GATCGACGGTACGCAAC | Fw sequencing primer pJULgldA |
| seq 2 | GGGTGGTAAAGGATGTCG | Rv sequencing primer pJULgldA |
| seq 3 | GCAACCTGGTTTGAAGC | Fw sequencing primer pJULgldA |
| seq 4 | GTGTTCGCTTCAATCACG | Rv sequencing primer pJULgldA |
For oligonucleotides, the prerequisite homologous region for Gibson assembly (underlined) and restriction sites (bold) are given and refer to the respective features or enzymes named in parenthesis