Facile assembly and fluorescence-based screening method for heterologous expression of biosynthetic pathways in fungi
Metabolic Engineering May 26, 2018 Sandra Hoefgen, Jun Lin, Janis Fricke et al. 113 citations
Expressing multiple genes from a biosynthetic pathway in eukaryotic hosts is challenging because each gene typically requires its own regulatory elements. A new vector system overcomes this by arranging genes as a single polycistron, using a picornavirus-inspired 'stop-carry on' mechanism so that all genes are controlled by one promoter. A split fluorescent reporter gene enables easy selection of transformed colonies. The method successfully produced high yields of the mushroom alkaloid psilocybin by expressing the entire biosynthetic gene cluster in the mould Aspergillus nidulans.