ArticlePloS one2013
Heterologous reconstitution of the intact geodin gene cluster in Aspergillus nidulans through a simple and versatile PCR based approach.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 44 papers.
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44 citing papers in PubMed, 100 citations in OpenAlex.
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- Halogenation in Fungi: What Do We Know and What Remains to Be Discovered?Molecules (Basel, Switzerland) · 2022Review
- Genome Mining and Analysis of PKS Genes inJournal of fungi (Basel, Switzerland) · 2022Article
- Increasing Lovastatin Production by Re-routing the Precursors Flow of Aspergillus terreus via Metabolic Engineering.Molecular biotechnology · 2022Article
- Transcriptional Activation of Biosynthetic Gene Clusters in Filamentous Fungi.Frontiers in bioengineering and biotechnology · 2022Review
- Comprehensive Guide to Extracting and Expressing Fungal Secondary Metabolites with Aspergillus fumigatus as a Case Study.Current protocols · 2021Article
- Halogenases: a palette of emerging opportunities for synthetic biology-synthetic chemistry and C-H functionalisation.Chemical Society reviews · 2021Review
- A CRISPR/Cas9 Cleavage System for Capturing Fungal Secondary Metabolite Gene Clusters.Journal of microbiology and biotechnology · 2021Article
- Gene mining and efficient biosynthesis of a fungal peptidyl alkaloid.Chinese herbal medicines · 2021Article
- In the fungus where it happens: History and future propelling Aspergillus nidulans as the archetype of natural products research.Fungal genetics and biology : FG & B · 2020Article
Corrections and comments
- Erratum issuedAnyaogu, Dianna Chinyere [corrected to Anyaogu, Diana Chinyere]
Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Fungal natural products are a rich resource for bioactive molecules. To fully exploit this potential it is necessary to link genes to metabolites. Genetic information for numerous putative biosynthetic pathways has become available in recent years through genome sequencing. However, the lack of solid methodology for genetic manipulation of most species severely hampers pathway characterization. Here we present a simple PCR based approach for heterologous reconstitution of intact gene clusters. Specifically, the putative gene cluster responsible for geodin production from Aspergillus terreus was transferred in a two step procedure to an expression platform in A. nidulans. The individual cluster fragments were generated by PCR and assembled via efficient USER fusion prior to transformation and integration via re-iterative gene targeting. A total of 13 open reading frames contained in 25 kb of DNA were successfully transferred between the two species enabling geodin synthesis in A. nidulans. Subsequently, functions of three genes in the cluster were validated by genetic and chemical analyses. Specifically, ATEG_08451 (gedC) encodes a polyketide synthase, ATEG_08453 (gedR) encodes a transcription factor responsible for activation of the geodin gene cluster and ATEG_08460 (gedL) encodes a halogenase that catalyzes conversion of sulochrin to dihydrogeodin. We expect that our approach for transferring intact biosynthetic pathways to a fungus with a well developed genetic toolbox will be instrumental in characterizing the many exciting pathways for secondary metabolite production that are currently being uncovered by the fungal genome sequencing projects.
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