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Direct cloning and heterologous expression of the salinomycin biosynthetic gene cluster from Streptomyces albus DSM41398 in Streptomyces coelicolor A3(2).
Yin, Jia Hoffmann, Michael Bian, Xiaoying Tu, Qiang Yan, Fu Xia, Liqiu Ding, Xuezhi Stewart, A Francis Müller, Rolf Fu, Jun
Yin, Jia
Hoffmann, Michael
Bian, Xiaoying
Tu, Qiang
Yan, Fu
Xia, Liqiu
Ding, Xuezhi
Stewart, A Francis
Müller, Rolf
Fu, Jun
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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.
Citation
Direct cloning and heterologous expression of the salinomycin biosynthetic gene cluster from Streptomyces albus DSM41398 in Streptomyces coelicolor A3(2). 2015, 5:15081 Sci Rep
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en
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2045-2322