Engineering a Streptomyces coelicolor biosynthesis pathway into Escherichia coli for high yield triglyceride production

Santiago Comba; Martín Sabatini; Simón Menendez-Bravo; Ana Arabolaza; Hugo Gramajo

doi:10.1186/s13068-014-0172-0

Table 1 Strains and plasmids

From: Engineering a Streptomyces coelicolor biosynthesis pathway into Escherichia coli for high yield triglyceride production

Strain or plasmid	Description	Reference
Strains
DH5α	E. coli K12 F^-lacU169 ( Φ80lacZ∆M15) endA1 recA1 hsdR17 deoR supE44 thi-1 l2 gyrA96 relA1	[40]
BL21 (DE3)	E. coli F^-ompT gal dcm lon hsdS_B(r_B^-m_B^-) λ(DE3)	Novagen
MPS10	BL21 (DE3) ∆dgkA::Km; Km^R	This study
MPS11	BL21 (DE3) ∆dgkA	This study
MPS12	BL21 (DE3) ∆dgkA ∆fadE::Km; Km^R	This study
MPS13	BL21 (DE3) ∆dgkA ∆fadE	This study
MPS14	BL21 (DE3) ∆dgkA ∆fadE ∆araBAD::Km; Km^R	This study
MPS15	BL21 (DE3) ∆dgkA ∆fadE ∆araBAD	This study
Plasmids
pET28a	Vector for expression of N-terminal His-tagged proteins under the strong T7 promoter; Km^R	Novagen
pBAD33	Vector for recombinant protein expression under the control of the P_BAD promoter; Cm^R	[41]
pKD13	Template plasmid for amplification of the FRT-flanked kanamycin cassette; Ap^R Km^R	[39]
pKD46	Temperature-sensitive replication plasmid for Red recombinase expression; Ap^R	[39]
pCP20	Temperature-sensitive replication plasmid for thermal induction of FLP synthesis; Cm^R Ap^R	[42]
pCR®-BluntII-TOPO	Vector used for cloning of blunt PCR products; Km^R	Invitrogen
pBAD0958	pBAD33 carrying the SCO0958^His gene under the control of P_BAD promoter; Cm^R	[15]
pET28-LPPβ	pET28 carrying the SCO1753^His gene under the control of T7 promoter; Km^R	[12]
pBAD-LPPβ	pBAD33 carrying the SCO1753^His gene under the control of P_BAD promoter; Cm^R	[12]
pTR257	pET28 carrying the SCO0958^His gene under the control of T7 promoter; Km^R	[15]
pET28-0958- LPPβ	pET28 carrying the SCO0958^His-SCO1753^His operon under the control of T7 promoter; Km^R	This study
pBAD-0958- LPPβ	pBAD33 carrying the SCO0958^His-SCO1753^His operon under the control of P_BAD promoter; Cm^R	This study
pET28-AtfA	pET28 carrying the atfA^His gene under the control of T7 promoter; Km^R	This study
pET28-FadD	pET28 carrying the fadD^His gene under the control of T7 promoter; Km^R	This study
pET28-0958- FadD	pET28 carrying the SCO0958^His-fadD^His operon under the control of T7 promoter; Km^R	This study
pCC01	pET28 carrying the accA2^His gene under the control of T7 promoter; Km^R	[43]
pET28-AccA2-AccE	pET28 carrying the accA2^His-accE^His operon under the control of T7 promoter; Km^R	This study
pET28-AccA2-AccE-AccB	pET28 carrying the accA2^His-accE^His-accB^His operon under the control of T7 promoter; Km^R	This study
pET28-0958-ACC	pET28 carrying the SCO0958^His-accA2^His-accE^His-accB^His operon under the control of T7 promoter; Km^R	This study
pET28-GpsA	pET28 carrying the gpsA^His gene under the control of T7 promoter; Km^R	This study
pET28-0958-GpsA	pET28 carrying the SCO0958^His-gpsA^His operon under the control of T7 promoter; Km^R	This study
pET28-FadR	pET28 carrying the fadR^His gene under the control of T7 promoter; Km^R	This study
pET28-0958-FadR	pET28 carrying the SCO0958^His-fadR^His operon under the control of T7 promoter; Km^R	This study
pET28-0958-0958	pET28 carrying the SCO0958^His-SCO0958^His operon under the control of T7 promoter; Km^R	This study

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