Table 3 E. coli strains, plasmids and primers used in this study
Strains | Relevant characteristics | Sources |
|---|---|---|
SZ470 | E. coli B, ΔfrdBC ΔldhA ΔackA ΔpflB ΔpdhR ::pflBp6-acEF-lpd ΔmgsA | [15] |
SZ85 | E. coli W3110, △focA-pflB △frdBC △adhE △ackA △ldhA::ldhL | [17] |
WL202 | SZ470, △adhE, lost anaerobic growth | This study |
WL203 | WL202, △ldhA::ldhL, regained anaerobic growth | This study |
WL204 | WL203, metabolically evolved in xylose with improved anaerobic growth | This study |
Plasmid | ||
pKD4 | FRT-kan-FRT cassette | [16] |
pKD46 | bla, red recombinase, temperature-dependent replication | [16] |
pFT-A | bla, flp, temperature-dependent replication | [20] |
Primers | ||
Clone ldhL-P1 | CCTATTATTTATGGCGGTGTCGTTT | This study |
Clone ldhL-P2 | CAGTTCGCTGACTGTAAGTTGTTGC | This study |
Delete adhE-P1 | ATGGCTGTTACTAATGTCGCTGAACTTAACGCAC | This study |
TCGTAGAGCGTGTGTAGGCTGGAGATGCTTC1) | ||
Delete adhE-P2 | TTAAGCGGATTTTTTCGCTTTTTTCTCAGCTTTAG | This study |
CCGGAGCAGCCATATGAATATCCTCCTTAG1) | ||
Verify △adhE-P1 | TGATGAAGGCTAATGCTG | This study |
Verify △adhE-P2 | CTTACGCCACCTGGAAGT | This study |
Verify insertion ldhL-P1 | GGTTCTAGTTACGCATTCG | This study |
Verify insertion ldhL-P2 | CTTCTTCTTTTCGTCATCG | This study |