Table 2.
E. coli strains, plasmids and primers used in this study
| Strains | Relevant characteristics | Sources |
|---|---|---|
| B | Wild type | ATCC |
| WL204 | E. coli B, ΔfrdBC ΔldhA ΔackA ΔpflB ΔpdhR ::pflBp6-acEF-lpd ΔmgsA ΔadhE, ΔldhA::ldhL, metabolically evolved in xylose with improved anaerobic growth | [30] |
| JH12 | WL204, ΔldhL::FRT-kan-FRT (chromosomal insertion of kan marker to replace ldhL gene), lost anaerobic cell growth, kanamycin positive | This study |
| JH13 | JH12, ΔFRT-kan-FRT::ldhA (chromosomal insertion of ldhA to replace kan marker), regaining anaerobic cell growth, kanamycin sensitive | This study |
| JH14 | JH13, ΔptsG::FRT-kan-FRT, slow glucose utilization due to disruption of the major glucose transporter system (PTS system) | This study |
| JH15 | JH14, metabolically evolved in glucose with improved glucose uptake (probably through alternative glucose transporter) | This study |
| Plasmids | ||
| pKD4 | FRT-kan-FRT cassette | [5] |
| pKD46 | bla, red recombinase, temperature-dependent replication | [5] |
| pFT-A | bla, flp, temperature-dependent replication | [22] |
| Primers | ||
| Deletion ldhL-P1 | TGTTTCGCTTCACCGGTCAGCTGTGTGTAGGCTGGAGATGCTTC a | This study |
| Deletion ldhL-P2 | TCGCTAATGGTGTTATCGAGTTAGCCATATGAATATCCTCCTTAG a | This study |
| Cloning ldhA-P1 | TGCAGCACGA AATCGCCCAG TTCAT | This study |
| Cloning ldhA-P2 | TGTGTGCATTACCCAACGGCAAACG | This study |
| Deletion ptsG-P1 | ATGTTTAAGAATGCATTTGCTAACCGTGTGTAGGCTGGAGATGCTTC a | This study |
| Deletion ptsG-P2 | TTAGTGGTTACGGATGTACTCATCCAAGCCATATGAATATCCTCCTTAG a | This study |
aThe underlined sequence is homologous to the flanked sequence of FRT-kan-FRT cassette in pKD4