Efficient targeted mutagenesis in Epichloë festucae using a split marker system

M Rahnama; N Forester; K G S U Ariyawansa; C R Voisey; L J Johnson; R D Johnson; D J Fleetwood

doi:10.1016/j.mimet.2016.12.017

Efficient targeted mutagenesis in Epichloë festucae using a split marker system

J Microbiol Methods. 2017 Mar:134:62-65. doi: 10.1016/j.mimet.2016.12.017. Epub 2016 Dec 22.

Authors

M Rahnama¹, N Forester², K G S U Ariyawansa², C R Voisey², L J Johnson², R D Johnson², D J Fleetwood³

Affiliations

¹ AgResearch, Grasslands Research Centre, Palmerston North, New Zealand; School of Biological Sciences, University of Auckland, New Zealand.
² AgResearch, Grasslands Research Centre, Palmerston North, New Zealand.
³ AgResearch, Grasslands Research Centre, Palmerston North, New Zealand; Biotelliga Ltd., Auckland, New Zealand. Electronic address: damien@biotelliga.com.

PMID: 28017692
DOI: 10.1016/j.mimet.2016.12.017

Abstract

A split-marker system for targeted gene deletion was developed for the model grass endophytic fungus Epichloë festucae. Compared to the conventional system that yields up to 25% homologous recombinants, the method resulted in 33-74% targeted deletions in E. festucae using as little as 1.5kb of targeting sequence.

Keywords: Endophyte; Epichloë; Functional analysis; Gene deletion; Split-marker.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cloning, Molecular / methods*
Epichloe / genetics*
Fungal Proteins / genetics*
Gene Deletion*
Genetic Vectors
Mutagenesis*
Phylogeny
Polymerase Chain Reaction
Recombination, Genetic

Substances

Fungal Proteins