Table 3 Bacterial strains and plasmids used in this study

Escherichia coli

W3110

F-, λ^-, IN(rrnD-rrnE)1, rph-1

KCTC

JW3973

F-, Δ(araD-araB)567, ΔlacZ4787(::rrnB-3),

Keio collection

λ^-, rph-1, ΔmetA780:: kan,

National Institute

Δ(rhaD-rhaB)568, hsdR514,

of Genetics, Japan

WE

JW3973 carrying the wild-type metA gene

[11]

K96

JW3973 carrying the metA gene

This study

with the Q96K substitution

L124

JW3973 carrying the metA gene

This study

with the I124L substitution

Y229

JW3973 carrying the metA gene

This study

with the I229Y substitution

Y247

JW3973 carrying the metA gene

This study

with the F247Y substitution

LY

JW3973 carrying the metA gene

This study

with the I124L and I229Y substitutions

LYD

JW3973 carrying the metA gene

This study

with the I124L, I229Y and N267D substitutions

WE(P^-)

WE (∆clpX-lon)::cat, ∆hslVU1172::tet

This study

L124(P^-)

L124 (∆clpX-lon)::cat, ∆hslVU1172::tet

This study

Y229(P^-)

Y229 (∆clpX-lon)::cat, ∆hslVU1172::tet

This study

WE∆dnaK

WE ∆dnaK::cat

This study

L124∆dnaK

L124∆dnaK::cat

This study

Y229∆dnaK

Y229∆dnaK::cat

This study

BL21(DE3)

F- ompT hsdS _B (r- _B m- _B ) gal dcm(DE3)

Novagen (Billerica, USA)

ME7970

∆hslVU1172::tet

National Institute

(KY2966)

of Genetics, Japan

Plasmids

pKD46

λ Red (gam bet exo) ara C rep101(Ts), Ap^r

[32]

pMetA

pACYC177 carrying the wild-type metA

[11]

under the natural P _metA promoter, Ap^r

pDnaK

pACYC177 carrying the dnaK gene

This study

under the natural P _dnaK promoter, Ap^r

pPP1

pACYC177 carrying the clpX-lon genes

This study

under the natural P _clpX promoter, Ap^r

pET22b/MetA

Contains the wild-type metA gene, Ap^r

[11]

pET22b/MetAL124

Contains the metA gene with I124L substitution, Ap^r

This study

pET22b/MetAY229

Contains the metA gene with I229Y substitution, Ap^r

This study