Construction and Diversification of Natural Product Biosynthetic Gene Clusters at High Efficiency and Accuracy.

ABSTRACT: Biosynthetic gene clusters (BGCs) encode the biosynthesis of natural products, which serve as the foundation for therapeutics such as antibiotics, anticancer agents, antifungals, and immunosuppressants. The vast majority of the BGCs remain uncharacterized due to lack of expression or inability to cultivate the native host, making refactoring and expression of BGCs in optimized hosts a prerequisite for genome-based drug discovery. Transformation-associated recombination (TAR) cloning and Gibson assembly are error prone due to the use of homologous recombination. Here, we present a BGC cloning and refactoring strategy based on a hierarchical Golden Gate Assembly (GGA), which enables systematic pathway engineering and mutagenesis with unprecedented accuracy and efficiency. We constructed the

SUBMITTER: Bai C 

PROVIDER: S-EPMC12645565 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature