Table S1. Bacterial strains and plasmids
Strain
Relevant genotype
Origin or construction
MG1655
Parental strain
Laboratory collection
DV1093
MVA+
(Vinella et al., 2009)
DV206
lacIpoZΔ(Mlu)
(Vinella et al., 2000)
DV1389
lacIpoZΔ(Mlu) PiscR::lacZ
DV1301
lacIpoZΔ(Mlu) PhmpA::lacZ
Lysogen of DV206 (This study)
DV1367
lacIpoZΔ(Mlu) PhmpA(short))::lacZ
Lysogen of DV206 (This study)
DV1479
lacIpoZΔ(Mlu) PiscR::lacZ MVA+
DV1307
lacIpoZΔ(Mlu) PhmpA::lacZ MVA+
DV1478
lacIpoZΔ(Mlu) PiscR::lacZ ∆iscR
Lysogen of DV206. As DV781
(Trotter et al., 2009)
DV1389 + P1/ DV1093, selection
LB Kan
DV1301 + P1/ DV1093, selection
LB Kan
DV1389 + P1/ JW2515, selection
LB Kan then cured with pCP20
DV1389 + P1/ JW1674 (Baba et
DV1471
al., 2006), selection LB Kan then
lacIpoZΔ(Mlu) PiscR::lacZ ∆sufA
cured with pCP20
DV1389 + P1/ DV595 (Vinella et
DV1473
al., 2009), selection LB Can then
lacIpoZΔ(Mlu) PiscR::lacZ ∆suf
cured with pCP20
DV1389 + P1/ DV698 (Vinella et
DV1474
al., 2009), selection LB Can then
lacIpoZΔ(Mlu) PiscR::lacZ ∆iscA
cured with pCP20
DV1389 + P1/ DV597 (Vinella et
DV1475
lacIpoZΔ(Mlu) PiscR::lacZ ∆iscUA
al., 2009), selection LB Can then
cured with pCP20
DV1480
DV1483
DV1135
DV1492
lacIpoZΔ(Mlu)
PiscR::lacZ
∆sufA DV1471 + P1/ DV1093, selection
MVA+
LB Kan
lacIpoZΔ(Mlu) PiscR::lacZ ∆iscA MVA+
lacIpoZΔ(Mlu)
PiscR::lacZ
MVA+
∆erpA::cat
lacIpoZΔ(Mlu) PiscR::lacZ
DV1474 + P1/ DV1093, selection
LB Kan
DV1479 + P1/ DV1094 (Vinella et
al., 2009), selection LB Can on LB
+ mevalonate plates in anaerobiosis
MVA+ ∆sufA
DV1471 + P1/ DV698 (Vinella et
1
al., 2009), selection LB Can on LB
∆iscA
+ mevalonate plates in anaerobiosis
DV1494
DV1336
lacIpoZΔ(Mlu) PiscR::lacZ MVA+ ∆sufA
∆iscA∆erpA::cat
DV1492 + P1/ DV1094 (Vinella et
al., 2009), selection LB Can on LB
+ mevalonate plates in anaerobiosis
DV1301 + P1/ JW4136, selection
lacIpoZΔ(Mlu) PhmpA::lacZ ∆nsrR
LB Kan then cured with pCP20
DV1301 + P1/ JW1674 (Baba et
DV1302
al., 2006), selection LB Kan then
lacIpoZΔ(Mlu) PhmpA::lacZ ∆sufA
cured with pCP20
DV1301 + P1/ DV595 (Vinella et
DV1306
al., 2009), selection LB Can then
lacIpoZΔ(Mlu) PhmpA::lacZ ∆suf
cured with pCP20
DV1301 + P1/ DV698 (Vinella et
DV1563
al., 2009), selection LB Can then
lacIpoZΔ(Mlu) PhmpA::lacZ ∆iscA
cured with pCP20
DV1301 + P1/ DV597 (Vinella et
DV1564
lacIpoZΔ(Mlu) PhmpA::lacZ ∆iscUA
al., 2009), selection LB Can then
cured with pCP20
DV1308
DV1570
DV1326
DV1576
DV1583
pBAD24
pBADI
pLAI-A
lacIpoZΔ(Mlu)
PhmpA::lacZ
∆sufA DV1302 + P1/ DV1093, selection
MVA+
lacIpoZΔ(Mlu)
LB Kan
PhmpA::lacZ
∆iscA DV1563 + P1/ DV1093, selection
MVA+
LB Kan
lacIpoZΔ(Mlu)PhmpA::lacZ
MVA+
∆erpA::cat
lacIpoZΔ(Mlu)
PhmpA::lacZ
MVA+
PhmpA::lacZ
MVA+
Cloning vector, AmpR
Cloning vector, pBAD24 derivative
Para::iscA, AmpR
DV1308 + P1/ DV698 (Vinella et
al., 2009), selection LB Can on LB
+ mevalonate plates in anaerobiosis
∆iscA ∆erpA::cat ∆sufA
(AmpR)
al., 2009), selection LB Can on LB
+ mevalonate plates in anaerobiosis
∆sufA ∆iscA
lacIpoZΔ(Mlu)
DV1307 + P1/ DV1094(Vinella et
DV1570 + P1/ DV1094 (Vinella et
al., 2009), selection LB Can on LB
+ mevalonate plates in anaerobiosis
(Guzman et al., 1995)
(Py et al., 1999)
(Vinella et al., 2009)
2
pLAS-A
Para::sufA, AmpR
(Vinella et al., 2009)
pLAE-A
Para::erpA, AmpR
(Vinella et al., 2009)
pLAOS
Para::sufABCDSE, AmpR
This study
pLAOa
Para::sufBCDSE, AmpR
This study
pLAOHyb
Para::iscAsufBCDSE, AmpR
This study
Red helper plasmid, pINT-ts derivative
pKD46
containing
araC-ParaB
and
γβexo, (Datsenko and Wanner, 2000)
AmpR
pET22(b)+
pET::nsrR
pET::iscR
Cloning and expression vector, AmpR Invitrogen
pET22 derivative encoding NsrR-His,
AmpR
This study
pET22 derivative encoding IscR-His,
AmpR
This study
3
Table S2. Sequence (5’ to 3’) of the oligonucleotides used in this work.
Primers
5SufB
3SufB
EcoRI-sufA
XhoI-sufA
NdeI-iscR
XhoI-iscR
NdeI-nsrR
XhoI-nsrR
5TFhmp
3TFhmp
Sequence
Cloning of sufB and sufA
GCATGAATTCATGTCTCGTAATACTGAAGCAAC
CCTCTCGAGTTATCCGACGCTGTGTTCAAGACTGATGGC
CTTCGAATTCATGGACATGCATTCAGGAACCTTTAAC
GTTCTCGAGCTATACCCCAAAGCTTTCGCCACAGC
Cloning of iscR and nsrR(in pET22)
CGCCATATGAGACTGACATCTAAAGGG
GTTCTCGAGAGCGCGTAACTTAACGTCGATCGC
TATACATATGCAGTTAACGAGTTTCAC
CCGCTCGAGCTCCACCAGCAATAATTTATAAAGCGG
Construction of the PhmpA::lacZ gene fusion
GCGGAATTCGTTGGACAACGGCGAACAG
Construction of the Phmp::lacZ gene fusion
GACGGGATCCCCGGTTTGAGCGTCAAGCATATGGTC
boiteNsr/hmpA
4
5