title: Development of the gateway recycling cloning system for multiple linking of expression cassettes in a defined order, and direction on gateway compatible binary vectors.
abstract: We developed the Gateway recycling cloning system, which allows multiple linking of expression cassettes by multiple rounds of the Gateway LR reaction. Employing this system, the recycling donor vector pRED419 was subjected to the first LR reaction with an attR1-attR2 type destination vector. Then conversion vector pCON was subjected to an LR reaction to restore the attR1-attR2 site on the destination vector for the next cloning cycle. By repetition of these two simple steps, we linked four expression cassettes of a reporter gene in Gateway binary vector pGWB1, introduced the constructs into tobacco BY-2 cells, and observed the expression of transgenes.
authors: Department of Sustainable Resource Science, Graduate School of Bioresources, Mie University, Tsu, Japan.
authors: Tetsuya
authors: T
authors: Kimura
authors: Akihide
authors: A
authors: Nakao
authors: Sachiko
authors: S
authors: Murata
authors: Yasuyuki
authors: Y
authors: Kobayashi
authors: Yuji
authors: Y
authors: Tanaka
authors: Kenta
authors: K
authors: Shibahara
authors: Tetsu
authors: T
authors: Kawazu
authors: Tsuyoshi
authors: T
authors: Nakagawa
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D015870:Gene Expression; D017930:Genes, Reporter; D005818:Genetic Engineering; D005822:Genetic Vectors; D059828:Plant Cells / Q000378:metabolism; D011995:Recombination, Genetic; D019076:Transgenes; D062789:Tobacco Products