| Literature DB >> 35524045 |
Mikkel Rank Nielsen1, Jens Laurids Sørensen2.
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.Entities:
Keywords: Chromosomal integration; Fungi; Fusarium; Gene cluster; Heterologous expression; Homologous recombination; Protoplast; Protoplast-mediated transformation; TAR
Mesh:
Year: 2022 PMID: 35524045 DOI: 10.1007/978-1-0716-2273-5_4
Source DB: PubMed Journal: Methods Mol Biol ISSN: 1064-3745