Acetate fluxes in Escherichia coli are determined by the thermodynamic control of the Pta-AckA pathway
Brice Enjalbert†, Pierre Millard†, Mickael Dinclaux, Jean-Charles Portais*, and Fabien Letisse
LISBP, Université de Toulouse, CNRS, INRA, INSA, Toulouse, France
†
these authors contributed equally to this manuscript *
[email protected]
1
Supplementary Tables
Supplementary Table S1. Primers used in this work and their function. Name H1 acs H2 acs
Sequence (5’ to 3’)
Function
AACGCTTATGCCACATATTATTAACATCCTA CAAGGAGAACAAAAGCATG GTTACCGACTCGCATCGGGCAATTGTGGGTT ACGATGGCATCGCGATAGC
External, 5’ of the acs gene External, 3’ of the acs gene Intern, toward 5’ of the acs gene Intern, toward 3’ of the acs gene External, 5’ of the ackA gene External, 3’ of the ackA gene Intern, toward 5’ of the ackA gene Intern, toward 3’ of the ackA gene External, 5’ of the poxB gene External, 3’ of the poxB gene Intern, toward 5’ of the poxB gene Intern, toward 3’ of the poxB gene Intern, toward 5’ of the kanamycine gene Intern, toward 3’ of the kanamycine gene
100 acs 5’
GGGAAAATTGACTGGCAGGA
100 acs 3’
GGATCTTCGGCGTTCATCTC
H1 ackA H2 ackA
TGGCTCCCTGACGTTTTTTTAGCCACGTATC AATTATAGGTACTTCCATG GCACCGCCAGCTGAGCTGGCGGTGTGAAAT CAGGCAGTCAGGCGGCTCGC
100 ackA 5’
TCTTCCACCTGCACGACACC
100 ackA 3’
TCGCTGGTCACTTCGGTCAG
H1 poxB H2 poxB
GATGAACTAAACTTGTTACCGTTATCACAT TCAGGAGATGGAGAACCATG CCTTATTATGACGGGAAATGCCACCCTTTT TACCTTAGCCAGTTTGTTTT
100 poxB 5’
TCGTGTTACCAGGCGACGTG
100 poxB 3’
CTTCTTCCGGCGTCACGACT
K1
CAGTCATAGCCGAATAGCCT
K2
CGGTGCCCTGAATGAACTGC
Supplementary Table S2. Strains used in this work. Name MG1655 BW25113 Δacs BW25113 ΔackA BW25113 ΔpoxB MG1655 Δacs MG1655 ΔackA MG1655 ΔpoxB
Genotype F- λ- ilvG- rfb-50 rph-1 rrnB3 ΔlacZ4787 hsdR514Δ(araBAD)567 Δ(rhaBAD)568 rph-1 Δacs::kan rrnB3 ΔlacZ4787 hsdR514Δ(araBAD)567 Δ(rhaBAD)568 rph-1 ΔackA::kan rrnB3 ΔlacZ4787 hsdR514Δ(araBAD)567 Δ(rhaBAD)568 rph-1 ΔpoxB::kan
Origin Blattner et al., 1997
F- λ- ilvG- rfb-50 rph-1 Δacs::kan
This work
F- λ- ilvG- rfb-50 rph-1 ΔackA::kan
This work
F- λ- ilvG- rfb-50 rph-1 ΔpoxB::kan
This work
2
Baba et al., 2006 Baba et al., 2006 Baba et al., 2006
