ArticleMethods in molecular biology (Clifton, N.J.)2022
Rapid Combinatorial Coexpression of Biosynthetic Genes by Transient Expression in the Plant Host Nicotiana benthamiana.
Article in Methods in molecular biology (Clifton, N.J.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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8 citing papers in PubMed, 15 citations in OpenAlex.
- Nicotine biosynthesis is completed by cryptic activating glucosylation.Nature communications · 2026Article
- Chemoenzymatic Synthesis with Plant Oxidases and Metabolic Engineering Enable Rapid Access to Rare Gibberellins.Journal of the American Chemical Society · 2026Article
- Phylogenomics and metabolic engineering reveal a conserved gene cluster in Solanaceae plants for withanolide biosynthesis.Nature communications · 2025Article
- Expansion of the Stereochemical Space of Triterpenes by Mining Noncanonical Oxidosqualene Cyclases Across the Diversity of Green Plants.Journal of the American Chemical Society · 2025Article
- Multiplicity of theACS synthetic biology · 2023Article
- Post-Cyclization Skeletal Rearrangements in Plant Triterpenoid Biosynthesis by a Pair of Branchpoint Isomerases.Journal of the American Chemical Society · 2023Article
- Cytochrome P450 monooxygenase-mediated tailoring of triterpenoids and steroids in plants.Beilstein journal of organic chemistry · 2022Review
- Identification of early quassinoid biosynthesis in the invasive tree of heaven (Frontiers in plant science · 2022Article
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2 authors at 1 institution in 1 country.
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Abstract
Metabolic engineering of heterologous hosts requires coexpression of multiple genes, often more than ten for a single pathway. Traditional approaches to create genetic constructs for this purpose are highly inflexible and suffer from very low throughput. In this book chapter, we describe a powerful method to overcome this bottleneck, namely, combinatorial co-expression in the Australian tobacco plant Nicotiana benthamiana. This system is based on Agrobacterium tumefaciens-mediated transient gene expression, often called agroinfiltration. Herein, instead of creating complex multigenic constructs, coexpression is achieved by simply mixing different plasmid-bearing Agrobacterium strains without the need for different selection markers. We present a practical guide for coexpressing multiple biosynthetic genes followed by GC-MS analysis, using production of the plant triterpene β-amyrin as an example. Our chapter provides a guideline to harness the potential of this versatile expression system in the natural product community for studying and engineering metabolic pathways.
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