Plasmids for increased efficiency of vector construction and genetic engineering in filamentous fungi

•We created a series of Gateway-enabled plasmids for use in filamentous fungi.•These plasmids can be used in gene deletion, gene integration, and high-copy gene expression.•The plasmids contain a variety of nutritional and drug-resistance markers that are functional across multiple fungal groups.

Fungal species are continuously being studied to not only understand disease in humans and plants but also to identify novel antibiotics and other metabolites of industrial importance. Genetic manipulations, such as gene deletion, gene complementation, and gene over-expression, are common techniques to investigate fungal gene functions. Although advances in transformation efficiency and promoter usage have improved genetic studies, some basic steps in vector construction are still laborious and time-consuming. Gateway cloning technology solves this problem by increasing the efficiency of vector construction through the use of λ phage integrase proteins and att recombination sites. We developed a series of Gateway-compatible vectors for use in genetic studies in a range of fungal species. They contain nutritional and drug-resistance markers and can be utilized to manipulate different filamentous fungal genomes.