title: Direct cloning and heterologous expression of the salinomycin biosynthetic gene cluster from Streptomyces albus DSM41398 in Streptomyces coelicolor A3(2).
abstract: Linear plus linear homologous recombination-mediated recombineering (LLHR) is ideal for obtaining natural product biosynthetic gene clusters from pre-digested bacterial genomic DNA in one or two steps of recombineering. The natural product salinomycin has a potent and selective activity against cancer stem cells and is therefore a potential anti-cancer drug. Herein, we separately isolated three fragments of the salinomycin gene cluster (salO-orf18) from Streptomyces albus (S. albus) DSM41398 using LLHR and assembled them into intact gene cluster (106 kb) by Red/ET and expressed it in the heterologous host Streptomyces coelicolor (S. coelicolor) A3(2). We are the first to report a large genomic region from a Gram-positive strain has been cloned using LLHR. The successful reconstitution and heterologous expression of the salinomycin gene cluster offer an attractive system for studying the function of the individual genes and identifying novel and potential analogues of complex natural products in the recipient strain.
authors.affiliation: Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shanda Nanlu 27, Jinan, 250100, People's Republic of China.|Department of Genomics, Dresden University of Technology, BioInnovations-Zentrum, Tatzberg 47-51, Dresden, 01307, Germany.|Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research and Department of Pharmaceutical Biotechnology, Saarland University, PO Box 151150, Saarbrücken, 66041, Germany.|Hunan Provincial Key Laboratory for Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, 410081, People's Republic of China.
authors.forename: Jia
authors.initials: J
authors.lastname: Yin
authors.affiliation: Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research and Department of Pharmaceutical Biotechnology, Saarland University, PO Box 151150, Saarbrücken, 66041, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Hoffmann
authors.affiliation: Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shanda Nanlu 27, Jinan, 250100, People's Republic of China.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Bian
authors.affiliation: Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shanda Nanlu 27, Jinan, 250100, People's Republic of China.|Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research and Department of Pharmaceutical Biotechnology, Saarland University, PO Box 151150, Saarbrücken, 66041, Germany.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Tu
authors.affiliation: Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research and Department of Pharmaceutical Biotechnology, Saarland University, PO Box 151150, Saarbrücken, 66041, Germany.
authors.forename: Fu
authors.initials: F
authors.lastname: Yan
authors.affiliation: Hunan Provincial Key Laboratory for Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, 410081, People's Republic of China.
authors.forename: Liqiu
authors.initials: L
authors.lastname: Xia
authors.affiliation: Hunan Provincial Key Laboratory for Microbial Molecular Biology-State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, 410081, People's Republic of China.
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Ding
authors.affiliation: Department of Genomics, Dresden University of Technology, BioInnovations-Zentrum, Tatzberg 47-51, Dresden, 01307, Germany.
authors.forename: A Francis
authors.initials: AF
authors.lastname: Stewart
authors.affiliation: Helmholtz Institute for Pharmaceutical Research, Helmholtz Centre for Infection Research and Department of Pharmaceutical Biotechnology, Saarland University, PO Box 151150, Saarbrücken, 66041, Germany.
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
authors.affiliation: Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shanda Nanlu 27, Jinan, 250100, People's Republic of China.|Department of Genomics, Dresden University of Technology, BioInnovations-Zentrum, Tatzberg 47-51, Dresden, 01307, Germany.
authors.forename: Jun
authors.initials: J
authors.lastname: Fu
authors.affiliation: Shandong University-Helmholtz Institute of Biotechnology, State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shanda Nanlu 27, Jinan, 250100, People's Republic of China.
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D022202:Chromosomes, Artificial, Bacterial; D003001:Cloning, Molecular; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors / Q000235:genetics; D005810:Multigene Family; D011714:Pyrans / Q000378:metabolism*; D048372:Streptomyces coelicolor / Q000235:genetics* / Q000378:metabolism*; D000095662:Polyether Polyketides
chemical_list: D011714:Pyrans; C010327:salinomycin; D000095662:Polyether Polyketides