Direct cloning and heterologous expression of the salinomycin biosynthetic gene cluster from Streptomyces albus DSM41398 in Streptomyces coelicolor A3(2)

• DOI: 10.1038/srep15081
• Publication Date: 2015-10-13T09:25:49Z
• Authors: Jia Yin; Michael Hoffmann; Xiaoying Bian; Qiang Tu; Fu Yan; Liqiu Xia; Xuezhi Ding; A. Francis Stewart; Rolf Müller; Jun Fu; Youming Zhang
• Keywords: Streptomyces albus; Salinomycin; Recombineering; Gene cluster; Heterologous expression; Cloning (programming); Heterologous; genomic DNA

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 salinomycin has a potent and selective activity against cancer stem cells therefore potential anti-cancer drug. Herein, we separately isolated three fragments the cluster ( sal O -orf 18) Streptomyces albus S. ) DSM41398 using LLHR assembled them into intact (106 kb) by Red/ET expressed it heterologous host coelicolor A3(2). We are first to report large region Gram-positive strain been cloned LLHR. successful reconstitution expression offer an attractive system studying function individual genes identifying novel analogues complex products recipient strain.