Related Experiment Video
Updated: May 14, 2026
Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit
Published on: October 31, 2019
Development of the gateway recycling cloning system for multiple linking of expression cassettes in a defined order,
Tetsuya Kimura1, Akihide Nakao, Sachiko Murata
1Department of Sustainable Resource Science, Graduate School of Bioresources, Mie University, Tsu, Japan.
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.
