Fungal gene knockout with Agrobacterium tumefaciens using Gibson assembly v1

• DOI: 10.17504/protocols.io.q26g7r3rqvwz/v1
• Publication Date: 2024-08-14T00:45:23Z
• Authors: Johannes Wolfram ...
• Keywords: Gene knockout

This method uses Agrobacterium tumefaciens to transfer a piece of TDNA containing resistance gene (hph for hygromycin in this particular case) flanked by homologous regions the you want knock out into fungal spore. The process is broken down following steps: Assembly knockout construct via Gibson assembly Transformation E.coli Plasmid harvest and transformation spores Verifying