Fungal Artificial Chromosome (FAC): A Novel Technology to Translate the Whole Genome Sequence Information into the Secondary Metabolome in Filamentous Fungi

With thousands of fungal genomes being sequenced, each genome containing up to 100 secondary metabolite (SM) clusters 30 - 80 kb in size, breakthrough techniques are needed to characterize this SM wealth. We have developed a novel system-level methodology for unbiased cloning of intact SM clusters from each of 6 fungal genomes for one-step transformation and expression in a model host. All intact SM clusters from 6 species of Aspergillus, Fusarium and Penicillin were individually captured in self-replicating fungal artificial chromosomes (FAC) containing both E. coli F replicon and an Aspergillus autonomously replicating sequence. Candidate FACs were successfully shuttled between E. coli and the heterologous expression host A. nidulans. We will present the new small molecule analysis pipeline and the discovery of fungal SM biosynthetic machineries.