Calibrating Transcriptional Activity Using Constitutive Synthetic Promoters in Mutants for Global Regulators in Escherichia coli

Table 1

Strains, plasmids and primers used in this study.

Strains, plasmids, and primers	Description	Reference
Strains
E. coli DH5α	F^− endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG purB20 φ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK^−mK^+), λ^−.	[12]
E. coli DH10B	mcrA Δmrr-hsdRMS-mcrBC) φ 80lacZΔM15 ΔlacX74 recA1 araD139 Δ (ara-leu)7697 galU galK rpsL endA1 nupG Δdcm.	[12]
E. coli BW25113	lacI^+rrnBT14 ΔlacZWJ16 hsdR514 ΔaraBADAH33 ΔrhaBADLD78 rph-1	[12]
	Δ(araB–D)567 Δ(rhaD–B)568 ΔlacZ4787(::rrnB-3) hsdR514 rph-1.
E. coli JW1702	E. coli BW25113 Δihf mutant	[12]
E. coli JW3229	E. coli BW25113 Δfis mutant	[12]
Plasmids
pMR1	Cm^R, ori p15a. GFPlva promoter probe vector	[14]
pMR1-Pj113	pMR1 with Pj113 cloned as EcoRI/BamHI fragment	This work
pMR1-Pj114	pMR1 with Pj114 cloned as EcoRI/BamHI fragment	This work
pMR1-Pj106	pMR1 with Pj106 cloned as EcoRI/BamHI fragment	This work
pMR1-Pj100	pMR1 with Pj100 cloned as EcoRI/BamHI fragment	This work
pMR1-Plac	pMR1 with Plac promoter cloned as EcoRI/BamHI fragment	[28]
pGLR2	Km^R, oriT, ori RK2. SEVA-based vector with dual GFP-lux reporter	[15]
pGLR2-Pj113	pGLR2 with Pj113 cloned as EcoRI/BamHI fragment	This work
pGLR2-Pj114	pGLR2 with Pj114 cloned as EcoRI/BamHI fragment	This work
pGLR2-Pj106	pGLR2 with Pj106 cloned as EcoRI/BamHI fragment	This work
pGLR2-Pj100	pGLR2 with Pj100 cloned as EcoRI/BamHI fragment	This work
pMR1-xylS-Pm	pMR1 with PxylS, xylS and Pm cloned as EcoRI/BamHI fragment	This work
Primers
Pj100-FW	GAATTCTTGACGGCTAGCTCAGTCCTAGG	This work
Pj100-RV	TACAGTGCTAGCAAGTGGATCCTTGCGATC	This work
Pj106-FW	GAATTCTTTACGGCTAGCTCAGTCCTAGGTA	This work
Pj106-RV	TAGTGCTAGCAAGTGGATCCTTGCGATC	This work
Pj114-FW	GAATTCTTTATGGCTAGCTCAGTCCTAGGT	This work
Pj114-RV	ACAATGCTAGCAAGTGGATCCTTGCGATC	This work
Pj113-FW	GAATTCCTGATGGCTAGCTCAGTCCTAGGG	This work
Pj113-RV	ATTATGCTAGCAAGTGGATCCTTGCGATC	This work
5_xylS_EcoRI	GAATTCTCAAGCCACTTCCTTTTTGCATTG	This work
3_Pm_BamHI	GGATCCATTATTGTTTCTGTTGCATAAAGCC	This work