title: "Cre/loxP plus BAC": a strategy for direct cloning of large DNA fragment and its applications in Photorhabdus luminescens and Agrobacterium tumefaciens.
abstract: Heterologous expression has been proven to be a valid strategy for elucidating the natural products produced by gene clusters uncovered by genome sequencing projects. Efforts have been made to efficiently clone gene clusters directly from genomic DNA and several approaches have been developed. Here, we present an alternative strategy based on the site-specific recombinase system Cre/loxP for direct cloning gene clusters. A type three secretion system (T3SS) gene cluster (~32 kb) from Photorhabdus luminescens TT01 and DNA fragment (~78 kb) containing the siderophore biosynthetic gene cluster from Agrobacterium tumefaciens C58 have been successfully cloned into pBeloBAC11 with "Cre/loxP plus BAC" strategy. Based on the fact that Cre/loxP system has successfully used for genomic engineering in a wide range of organisms, we believe that this strategy could be widely used for direct cloning of large DNA fragment.
authors.affiliation: Hunan Provincial Key Laboratory of 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.|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: Shengbiao
authors.initials: S
authors.lastname: Hu
authors.affiliation: Hunan Provincial Key Laboratory of 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: Zhengqiang
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Hunan Provincial Key Laboratory of 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: Xu
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Hunan Provincial Key Laboratory of 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: Guoyong
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Hunan Provincial Key Laboratory of 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: Yali
authors.initials: Y
authors.lastname: Xie
authors.affiliation: Hunan Provincial Key Laboratory of 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: Hunan Provincial Key Laboratory of 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: Xiangtao
authors.initials: X
authors.lastname: Mo
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: A Francis
authors.initials: AF
authors.lastname: Stewart
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: Hunan Provincial Key Laboratory of 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.|Department of Genomics, Dresden University of Technology, BioInnovations-Zentrum, Tatzberg 47-51, Dresden, 01307, Germany.
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Hunan Provincial Key Laboratory of 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
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D005822:Genetic Vectors; D016678:Genome / Q000235:genetics; D019426:Integrases / Q000235:genetics*; D020637:Photorhabdus / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic
chemical_list: C045073:Cre recombinase; D019426:Integrases