title: Genome engineering of Agrobacterium tumefaciens using the lambda Red recombination system.
abstract: Agrobacterium tumefaciens has been widely used as a tool for transgenesis in plants. The availability of its genome sequence should facilitate the directed engineering of improved properties; however, the current genome engineering options are laborious. Here, we investigated whether the lambda Red operon can be applied for recombineering of the A. tumefaciens genome. First, we built an expression plasmid for A. tumefaciens employing a tetracycline-inducible promoter to regulate the Red operon. This multicopy plasmid was then itself modified in A. tumefaciens to verify that the Red operon was functional. Then, we modified the endogenous A. tumefaciens tumor-inducing plasmid and the linear chromosome. These results extend recombineering technology to a new host and indicate a fast and convenient way to engineer the A. tumefaciens genome for functional genomics and strain improvements.
authors.affiliation: College of Life Science, Hunan Normal University, Hunan Provincial Key Laboratory of Microbial Molecular Biology, State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha, People's Republic of China.
authors.forename: Shengbiao
authors.initials: S
authors.lastname: Hu
authors.forename: Jun
authors.initials: J
authors.lastname: Fu
authors.forename: Fan
authors.initials: F
authors.lastname: Huang
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Ding
authors.forename: A Francis
authors.initials: AF
authors.lastname: Stewart
authors.forename: Liqiu
authors.initials: L
authors.lastname: Xia
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D010582:Bacteriophage lambda / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D016680:Genome, Bacterial; D009876:Operon; D010957:Plasmids; D011995:Recombination, Genetic