Investigating Fungal Biosynthetic Pathways Using Heterologous Gene Expression: Fusarium sp. as a Heterologous Host

Mikkel Rank Nielsen; Jens Laurids Sørensen

doi:10.1007/978-1-0716-2273-5_4

Investigating Fungal Biosynthetic Pathways Using Heterologous Gene Expression: Fusarium sp. as a Heterologous Host

Methods Mol Biol. 2022:2489:53-74. doi: 10.1007/978-1-0716-2273-5_4.

Authors

Mikkel Rank Nielsen¹, Jens Laurids Sørensen²

Affiliations

¹ Department of Chemistry and Bioscience, Aalborg University Esbjerg, Esbjerg, Denmark. mrn@bio.aau.dk.
² Department of Chemistry and Bioscience, Aalborg University Esbjerg, Esbjerg, Denmark.

PMID: 35524045
DOI: 10.1007/978-1-0716-2273-5_4

Abstract

Heterologous expression of uncharacterized biosynthetic gene clusters is a popular strategy for exploring the chemical potential of filamentous fungi. Here, we describe the process of PCR-amplifying fungal gene clusters and re-assembling them in a cloning vector via target-associated recombination in Saccharomyces cerevisiae . The gene cluster-carrying construct is validated and used to transform protoplasts of Fusarium graminearum , a well-studied host that is able to express the gene cluster. Chemical analysis of transformants expressing biosynthetic genes can lead to the detection and isolation of novel compounds, such as polyketides.

Keywords: Chromosomal integration; Fungi; Fusarium; Gene cluster; Heterologous expression; Homologous recombination; Protoplast; Protoplast-mediated transformation; TAR.

Publication types

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

MeSH terms

Biosynthetic Pathways* / genetics
Fusarium* / genetics
Gene Expression
Multigene Family
Saccharomyces cerevisiae / genetics