Table 1 Strains, plasmids and primers used in the study

Strains

E. coli B

F-

CGSC #2507

SSY01

E. coli B, ΔPDH-promoter::FRT-kan-FRT-ldhA gene promoter; promoter of pdh gene replaced with promoter of ldhA gene

This study

SSY02

E. coli B, ΔPDH-promoter::FRT-kan-FRT-frdA gene promoter

This study

SSY03

E. coli B, ΔPDH-promoter::FRT-kan-FRT-pflB gene promoter

This study

SSY04

E. coli B, ΔPDH-promoter::FRT-kan-FRT-adhE gene promoter

This study

SSY05

E. coli B, ΔPDH-promoter::FRT-kan-FRT-gapA gene promoter

This study

SSY06

SSY05 ΔldhA::FRT-kan-FRT; deletion mutant for ldhA gene in SSY05 host

This study

SSY07

SSY06 ΔfrdA::FRT-kan-FRT; deletion mutant for frdA gene in SSY06 host

This study

SSY08

SSY07 ΔackA::FRT-kan-FRT; deletion mutant for ackA gene in SSY07 host

This study

SSY09

SSY08 ΔpflB::FRT-kan-FRT; deletion mutant for pflB gene in SSY08 host

This study

Plasmids

pUC19

bla, cloning vector

pKD4

bla, FRT-kan-FRT

CGSC #7632

pKD46

bla, γ β exo (red recombinase), temperature-conditional replicon

CGSC #7739

pCP20

bla, flp, temperature-conditional replicon

CGSC #7629

pSSY01

FRT-kan-FRT sequence from pKD4 was cloned into pUC19 at Eco RI and Bam HI sites

This study

pSSY02

ldh A gene promoter from E. coli B was cloned into pSSY01 at Bam HI and Hind III site

This study

pSSY03

frd A gene promoter from E. coli B was cloned into pSSY01 at Bam HI and Hind III sites

This study

pSSY04

pfl B gene promoter from E. coli B was cloned into pSSY01 at Bam HI and Hind III sites

This study

pSSY05

adh E gene promoter from E. coli B was cloned into pSSY01 at Bam HI and Hind III sites

This study

pSSY06

gap A gene promoter from E. coli B was cloned into pSSY01 at Bam HI and Hind III sites

This study

pZSblank

P_LtetO1 expression vector, pSC101*origin, Cm^R

[11]

pZS*mcs

multiple cloning site derived from pET28a(+) cloned in pZSblank

This study

pZSack

ack gene cloned in pZS*mcs vector

This study

Primers

FRT-kan-FRT-F

GGAGAGAATTC GTGTAGGCTGGAGCTGCTTC

This study

FRT-kan-FRT-R

GGAGAGGATCC ATATGAATATCCTCCTTAG

This study

ldh A promoter-F

TCGGGATCC GCAAGCTGACAATCTCCC

This study

ldh A promoter-R

ACTCAAGCTT AAGACTTTCTCCAGTGATGTTG

This study

frd A promoter-F

TGCGGATCC ATCAAACAGCGGTGGGCAG

This study

frd A promoter-R

CCCAAGCTT GACATTCCTCCAGATTGTTT

This study

pfl B promoter-F

TCGGGATCC AACCATGCGAGTTACGGGCCTATAA

[8]

pfl B promoter-R

CCCAAGCTT GTGCCTGTGCCAGTGGTTGCTGTGA

This study

adh E promoter-F

CGCGGATCC CCGGATAATGTTAGCCATAA

This study

adh E promoter-R

CCCAAGCTT AATGCTCTCCTGATAATGTTA

This study

gap A promoter-F

CGCGGATCC GATTCTAACAAAACATTAACAC

This study

gap A promoter-R

CCCAAGCTT ATATTCCACCAGCTATTTGT

This study

H1

CTCCTTTCCTACGTAAAGTCTACATTTGTGCATAGTTACAACTTT GTGTAGGCTGGAGCTGCTTC

[8]

H2_ldhA

GCGAGTTTCGATCGGATCCACGTCATTTGGGAAACGTTCTGACAT AAGACTTTCTCCAGTGATGTTG

This study

H2_adhE

GCGAGTTTCGATCGGATCCACGTCATTTGGGAAACGTTCTGACAT AATGCTCTCCTGATAATGTT

This study

H2_gapA

GCGAGTTTCGATCGGATCCACGTCATTTGGGAAACGTTCTGACAT ATATTCCACCAGCTATTTGT

This study

H2_frdA

GCGAGTTTCGATCGGATCCACGTCATTTGGGAAACGTTCTGACAT GACATTCCTCCAGATTGTTT

This study

H2_pflB

GCGAGTTTCGATCGGATCCACGTCATTTGGGAAACGTTCTGACAT GTAACACCTACCTTCTGTTG

[8]

CTGTGATATAGAAGAC

v-PDH-F

TGCATGGTTGAAGATGAGTTG

This study

v-PDH-R

TGATGTAGTTGCTGATACCTG

This study

pET28mcs-F

CGGGATCC GAATTCGAGCTC

This study

pET28mcs-R

CTGACGCGT GTTAACAGCTTCCTTTCGGGCTTTG

This study

pZS-ack-F

GATCGGATCC ATGTCGAGTAAGTTAGTACTGGT

This study

pZS-ack-R

TCGAGTCGAC TCAGGCAGTCAGGCGGCTC

This study