Inactivation of PadR, the Repressor of the Phenolic Acid Stress Response, by Molecular Interaction with Usp1, a Universal Stress Protein from Lactobacillus plantarum, in Escherichia coli

Jérôme Gury; Hélène Seraut; Ngoc Phuong Tran; Lise Barthelmebs; Stéphanie Weidmann; Patrick Gervais; Jean-François Cavin

doi:10.1128/AEM.00774-09

TABLE 1.

Bacterial strains and plasmids

Strain or plasmid	Relevant characteristics^a	Source or reference
Strains
E. coli
TG1	supE hsdΔ5thi Δ(lac-proAB)F′ [traD36 proAB+ lacI^qlacZΔM15]	15
BL21(DE3) Star	F⁻ompT hsdS_B (r_B⁻ m_B⁻) gal dcm (DE3)	Invitrogen
TOP 10	F⁻mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 deoR araD139 Δ(ara-leu)7697 galU galK rpsL (Str^r) endA1 nupG	Invitrogen
L. plantarum
NC8 (wild type)	Wild type, gram-positive ubiquitous homolactic acid bacterium, plasmid-free strain	2
LPNC8ΔUSP1 (Δusp1)	L. plantarum NC8 strain with usp1 disrupted by double homologous recombination	This study (Fig. (Fig.33)
LPNC8ΔPADR (ΔpadR)	L. plantarum NC8 strain with padR disrupted by double homologous recombination	16; Fig. Fig.33
Plasmids
pTZ19R	Amp^r; ΔlacZ	Novagen
pET28a+	Kan^r; vector for overexpression of His-tagged proteins using the T7 bacteriophage promoter	Novagen
TOPO PCR 2.1	Amp^r Kan^r; ΔlacZ, vector for TA cloning	Invitrogen
pGID023	Shuttle vector for E. coli and L. plantarum; derivative of pJDC9 containing the pE194 replication functions; used as an unstable integration vector; Erm^r	18
pJPDC1	Em^r; pJDC9 containing the 2.3-kbp Sau3A fragment of L. plantarum corresponding to the locus with padA-padR genes	6; Fig. Fig.44
pLOCPAD	Amp^r Kan^r; TOPO vector containing the 2.212-kbp LOCPAD fragment with padA, padR, and usp1 PCR amplified with primers LPLOC1 and LPLOC2	This study (Fig. (Fig.44)
pTD14	Amp^r; pTZ19R containing the 311- and 385-bp DNA fragments PCR amplified with the primers LPDELU1-LPDELU2 and LPDELU3-LPDELU4, respectively	This study
pGΔUSP1	Em^r; pGID023 containing the 694-bp D14 fragment PCR amplified with the primers LPDELU1 and LPDELU4	This study
pED	pET28a+ with padA under its own promoter cloned into SphI restriction site	This study (Fig. (Fig.44)
pER	pET28a+ containing padR between NcoI and XhoI sites to produce PadR	16; Fig. Fig.44
pEU	pET28a+ containing usp1 between NcoI and XhoI sites to produce Usp1	This study (Fig. (Fig.44)
pEDR	pER with padA under its own promoter cloned into SphI restriction site	This study (Fig. (Fig.44)
pERU	pET28a+ with padR and usp1 cloned between NcoI and XhoI restriction sites	This study (Fig. (Fig.44)
pEDRU	pERU with padA under its own promoter cloned into SphI restriction site	This study (Fig. (Fig.44)

^aEm^r, erythromycin resistance; Amp^r, ampicillin resistance; Kan^r, kanamycin resistance.