Table 1 Bacterial strains, plasmids, and primers used in this study
From: Production of d-lactate from glucose using Klebsiella pneumoniae mutants
Strain, plasmid and primers | Description | Source |
|---|---|---|
Strains | ||
 E. coli DH5α | Cloning host | Lab collection |
 E. coli χ7213 | Host strain for pRE112, DAP auxotrophic strain | [14] |
 Q1188 | K. pneumoniae ATCC25955 | ATCC |
 Q2657 | K. pneumoniae ATCC25955 ΔadhE | This study |
 Q2666 | K. pneumoniae ATCC25955 ΔackA | This study |
 Q2699 | K. pneumoniae ATCC25955 ΔbudB | This study |
 Q2702 | K. pneumoniae ATCC25955 ΔbudB ΔackA ΔadhE | This study |
 Q2710 | K. pneumoniae ATCC25955 ΔbudB ΔadhE | This study |
 Q2743 | K. pneumoniae ATCC25955 ΔbudB ΔackA | This study |
Plasmids | ||
 pRE112 | Suicide vector, R6k origin, chloramphenicol resistant | [15] |
 pRE112-ΔbudB | Suicide vector for construction of ΔbudB mutant | This study |
 pRE112-ΔadhE | Suicide vector for construction of ΔadhE mutant | This study |
 pRE112-ΔackA | Suicide vector for construction of ΔackA mutant | This study |
Primers | Sequence (5′–3′) | Restriction enzymes |
|---|---|---|
ID-pRE112 | CAAGGCGACAAGGTGCTGATG | Â |
ΔadhE construction | ||
 1381 | GCTCTAGAATGGCTGTTACTAATATCGC | XbaI |
 1382 | CGACGCCGATAGCAGGTTTAC |  |
 1383 | GTAAACCTGCTATCGGCGTCGGTGCGTTCGGTGGTCTGGAT |  |
 1384 | GGGGTACCTCAGCCTTTACCGGAGCAAC | KpnI |
 1385 | ATAATGTCGAATCGAGCGAC |  |
 1386 | GCTTGTCGCGATGCTATCGC |  |
ΔackA construction | ||
 1387 | GCTCTAGAATGTCGAGTAAGTTAGTAC | XbaI |
 1388 | GCGTAGAGATAGGATTCTTC |  |
 1389 | GAAGAATCCTATCTCTACGCGAAGGCCTGGTGATGGGTAC |  |
 1390 | CGAGCTCTTATGCGGTCAGACGGCTGGC | SacI |
 1391 | ATCCTGCGCTACGCTAATGAC |  |
 1392 | CCTGCAGCTCGAATTATTGC |  |
ΔbudB construction | ||
 1448 | GCTCTAGAGTCAGTCAGCTGGAAGCTC | XbaI |
 1449 | ATAAGCTTCGCCACCTGGTC |  |
 1450 | GACCAGGTGGCGAAGCTTATCTGCGCATCGTTCGCGCCAT |  |
 1451 | CGAGCTCTTATCGCGATAATCTACCG | SacI |
 1452 | ATGGACAAACAGTATCCG |  |
 1453 | ACAGAATCTGACTCAGATG |  |
RT-qPCR | ||
 16S rRNA_F | AAGCGTTAATCGGAATTAC |  |
 16S rRNA_R | GCTACACCTGGAATTCTA |  |
 16S rRNA_Probe | (FAM)CTCTACAAGACTCTAGCCTGCCAG(Eclipse) |  |
 ldhA_F | GAAGCGGTATGTATCTTC |  |
 ldhA_R | CAGGGCGATATATTTCAC |  |
 ldhA_Probe | (FAM)CTTCAGCTCTTCCAGCACCG(ECLIPSE) |  |