SimReg1 is a master switch for biosynthesis and export of simocyclinone D8 and its precursors

Horbal, Liliya; Rebets, Yuriy; Rabyk, Mariya; Makitrynskyy, Roman; Luzhetskyy, Andriy; Fedorenko, Victor; Bechthold, Andreas

doi:10.1186/2191-0855-2-1

AMB Express

Table 1 Strains and plasmids

From: SimReg1 is a master switch for biosynthesis and export of simocyclinone D8 and its precursors

Bacterial strains and plasmids	Description	Source or reference
E. coli DH5α	supE44 ΔlacU169(φ80lacZΔM15)	Hanahan 1985
E. coli BL21 (DE3) pLysS	Host for the heterologous expression of His₆ -tagged simReg1	Novagen
E. coli ET12567/pUB307	hsdR17 recA1endA1gyrA96 thi-1 relA1 dam-13::Tn9(Cmr) dcm-6 hsd M; harbors conjugative plasmid pUB307; Cm^r, Km^r	Flett et al. 1997
S. antibioticus Tü6040	Simocyclinone D8 producing strain	Trefzer et al. 2002
S. antibioticus	Derivative of S. antibioticus Tü6040 with	This work
ΔsimReg1	disrupted simReg1 gene
S. antibioticus ΔsimReg1 × pSSimR1-1	ΔsimReg1 strain carrying plasmid with the intact simReg1 gene under its own promoter, used for complementation studies	This work
S. lividans 1326	Wild type	Hopwood et al. 1985
S. lividans × pSimD4script	Derivative of S. lividans 1326 carrying plasmid with gusA gene under the control of the putative promoter of the simD4 gene	This work
S. lividans × pSimD4script/pUWLsimReg1	Derivative of S. lividans 1326 carrying plasmid with gusA gene under the control of putative promoter of the simD4 gene and second plasmid with simReg1 gene under the control of P_ermE	This work
S. lividans ×pGUS	Derivative of S. lividans 1326 carrying plasmid with promoterless reporter gene gusA	This work
S. lividans × pGUS/pUWLsimReg1	Derivative of S. lividans 1326 carrying plasmid with promoterless reporter gene gusA and plasmid with simReg1 gene under the control of the P_ermEpromoter	This work
pBluescriptIIKS ⁺	General purpose cloning vector; Ap^r	MBI Fermentas
pUC19	General purpose cloning vector; Ap^r	MBI Fermentas
pSET152	E. coli/Streptomyces shuttle vector with φC31 integration system for streptomycetes; Am^r	Bierman et al. 1992
pKC1218E	pKC1218 derivative expression vector with P_ermEpromoter and SCP2* replicon; Am^r	Ostash et al. 2004
pHYG1	pLitmus38 containing hygromycin resistance cassette hyg	C. Olano Univ. de Oviedo, Spain
pKC1139	E. coli/Streptomyces shuttle vector with temperature sensitive replicon pSG5, Am^r	Muth et al. 1989
pUWL-oriT	pUWL-KS derivative harboring oriT from pSET152	Zelyas et al. 2009
pET21d	Vector for His-tagged protein expression	Novagen
5JH10	pUC plus simB3-D4 segment	Trefzer et al. 2002
pUCsimR1	pUC19 derivative containing simReg1 gene	This work
pUCsimR1-hyg	pUCsimR1 derivative with hyg cassette cloned into the simReg1 coding region	This work
pKCsimR1-hyg	pKC1139 derivative with cloned simReg1::hyg construction used for simReg1 gene inactivation	This work
pKCEsimR1	pKCE1218 derivative containing simReg1 gene under the control of P_ermE	Rebets et al. 2008
pSSimR1	pSET152 plus 2.3 kb simD4-X1 segment	This work
pSSimR1-1	pSET152 derivative containing simReg1 gene under the control of its own promoter	This work
pMA-simR1	plasmid containing synthetic codon-optimized simReg1 gene	Mr. Gene, Heidelberg
pETSR1c-15	pET21d derivative containing synthetic codon-optimized simReg1 gene	This work
pGUS	pSET152 derivative containing promoterless reporter gene gusA	Myronovskyi et al.2011
pSimD4script	derivative of pGUS harboring gusA reporter gene under the promoter of the simD4 gene	This work
pUWLsimReg1	derivative of pUWL containing gene simReg1 under the control of P_ermE	This work

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