ArticleMethods in enzymology2019
Direct cloning and heterologous expression of natural product biosynthetic gene clusters by transformation-associated recombination.
Article in Methods in enzymology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 33 papers.
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Who cites it
33 citing papers in PubMed.
- The PreKit platform: Cryptic gene clusters activation and high-titer compound production.Synthetic and systems biotechnology · 2026Article
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- Establishment of a protoplast transformation method for the oleaginous yeast Rhodotorula toruloides.FEMS yeast research · 2026Article
- Extracellular activity of a bacterial protease associated with reduced phage infectivity.PloS one · 2026Article
- Saskemycin, a potent and selective antimycobacterial agent targeting a unique site on the ribosome.Research square · 2025Article
- Discovery of acylsulfenic acid-featuring natural product sulfenicin and characterization of its biosynthesis.Nature chemistry · 2025Article
- Synthetic Biology in Natural Product Biosynthesis.Chemical reviews · 2025Review
- Enabling Access to Novel Bacterial Biosynthetic Potential From ONT Draft Genomic Data.Microbial biotechnology · 2025Article
- Genetic toolbox for Photorhabdus and Xenorhabdus: pSEVA based heterologous expression systems and CRISPR/Cpf1 based genome editing for rapid natural product profiling.Microbial cell factories · 2024Article
- CRISPR-aided genome engineering for secondary metabolite biosynthesis in Streptomyces.Journal of industrial microbiology & biotechnology · 2024Review
- Review
- Recent advances in the direct cloning of large natural product biosynthetic gene clusters.Engineering microbiology · 2023Review
- A unique dual acyltransferase system shared in the polyketide chain initiation of kidamycinone and rubiflavinone biosynthesis.Frontiers in microbiology · 2023Article
- Article
- Phylogeny-Informed Synthetic Biology Reveals Unprecedented Structural Novelty in Type V Glycopeptide Antibiotics.ACS central science · 2022Article
- A rapid and efficient strategy to identify and recover biosynthetic gene clusters from soil metagenomes.Applied microbiology and biotechnology · 2022Article
- Understanding and Manipulating Assembly Line Biosynthesis by Heterologous Expression in Streptomyces.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Renewed interests in the discovery of bioactive actinomycete metabolites driven by emerging technologies.Journal of applied microbiology · 2022Review
- Production of Epoxyketone Peptide-Based Proteasome Inhibitors byFrontiers in microbiology · 2022Article
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4 authors.
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Abstract
Heterologous expression of natural product biosynthetic gene clusters (BGCs) is a robust approach not only to decipher biosynthetic logic behind natural product (NP) biosynthesis, but also to discover new chemicals from uncharacterized BGCs. This approach largely relies on techniques used for cloning large BGCs into suitable expression vectors. Recently, several whole-pathway direct cloning approaches, including full-length RecE-mediated recombination in Escherichia coli, Cas9-assisted in vitro assembly, and transformation-associated recombination (TAR) in Saccharomyces cerevisiae, have been developed to accelerate BGC isolation. In this chapter, we summarize a protocol for TAR cloning large NP BGCs, detailing the process of choosing TAR plasmids, designing pathway-specific TAR vectors, generating yeast spheroplasts, performing yeast transformation, and heterologously expressing BGCs in various host strains. We believe that the established platforms can accelerate the process of discovering new NPs, understanding NP biosynthetic logic, and engineering biosynthetic pathways.
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