Related Experiment Video
Updated: Jul 16, 2026
Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing
Published on: July 7, 2020
Development of a new "GFP hop-on assay" system for insertion sequence transposition in Bacillus subtilis 168 using
Kiwamu Takahashi1, Taku Chibazakura, Yasuhiko Sekine
1Department of Bioscience, Tokyo University of Agriculture, Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan.
Abstract:
While most studies involving transposition have focused on analyzing the detailed mechanisms of transposition, the cellular conditions under which transposition occurs remain to be elucidated. In Escherichia coli, papillation assay is a powerful tool for transpositional analysis and the isolation of mutants affecting transposition. On the other hand, while our assay system based on the E. coli papillation assay can detect transpositional events in Bacillus subtilis 168, it is not suitable for quantitating transposition frequency because blue papillae on the transposant colonies of B. subtilis are not countable. We succeeded in developing a new "GFP hop-on assay" system that facilitates quantitative detection of the transposition of the FACS-optimized GFP mutant gene. Our assay system is a step forward in understanding the cellular conditions under which transposition occurs.
