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Table 2 E. coli strains, plasmids and primers used in this study

From: Enhancement of D-lactic acid production from a mixed glucose and xylose substrate by the Escherichia coli strain JH15 devoid of the glucose effect

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

  1. aThe underlined sequence is homologous to the flanked sequence of FRT-kan-FRT cassette in pKD4