ArticleMethods in molecular biology (Clifton, N.J.)2022
Heterologous Expression of Fungal Biosynthetic Pathways in Aspergillus nidulans Using Episomal Vectors.
Article in Methods in molecular biology (Clifton, N.J.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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5 citing papers in PubMed, 12 citations in OpenAlex.
- Protoplasting and Transformation Using CRISPR/Cas9 in Coccidioides.Current protocols · 2026Article
- Article
- Analyzing the Challenges and Opportunities Associated With Harnessing New Antibiotics From the Fungal Microbiome.MicrobiologyOpen · 2025Review
- Extracting Protoplasts from Filamentous Fungi Using Extralyse, An Enzyme Used in the Wine Industry.Current protocols · 2025Article
- Next-generation AMA1-based plasmids for enhanced heterologous expression in filamentous fungi.Microbial biotechnology · 2024Article
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Authors and funding
2 authors at 1 institution in 1 country.
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Abstract
Filamentous fungi produce a wide diversity of secondary metabolites, whose biosynthesis is encoded in biosynthetic gene clusters (BGCs). As novel BGCs are often found in fungal species that are genetically intractable or difficult to cultivate, heterologous expression is increasingly being used for compound discovery. In addition, heterologous expression is a useful strategy to elucidate the function of the genes within a BGC and shed light on their enzymatic mechanisms. Here, we describe a method for BGC elucidation using multi-marker AMA1-based pYFAC vectors for episomal expression in the fungal host Aspergillus nidulans. The pYFAC vectors have the advantage of high transformation efficiency and support high compound production. In addition, different pathway intermediates can be easily evaluated by testing different vector combinations. This protocol encompasses different AMA1-based strategies for BGC expression such as cloning of a BGC native sequence, promoter exchange or transcription factor overexpression. We also describe procedures for A. nidulans protoplasting, transformation, and small-scale culture analysis of strains containing AMA1 vectors.
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