Work

Protocol for connector-enabled multigene pathway assembly using Golden Gate Assembly and yeast recombination

Year: 2026

Type: article

Abstract: Here, we present a protocol for multigene pathway construction in Saccharomyces cerevisiae using a connector-enabled workflow that integrates Golden Gate Assembly with yeast homologous recombination. We provide procedures for assembling Level 1 transcriptional unit (TU) fragments using standardized connectors and directly co-transforming them into S. cerevisiae to form Level 2 multigene plasmids. Compared to conventional approaches, this protocol significantly reduces procedural complexity, minimizes dependence on operator expertise, and substantially accelerates workflow timelines. For complete details on the use and execution of this protocol, please refer to Seong et al. 1

Source: STAR Protocols

DOI: 10.1016/j.xpro.2026.104648

Authors: Min-Jun Seong, Jonghyeok Shin, Dae-Hee Lee

Institutions: Korea Research Institute of Bioscience and Biotechnology, Korea University of Science and Technology, Sejong University, Korea Advanced Institute of Science and Technology, Sungkyunkwan University

Language: English

Topic: Fungal and yeast genetics research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,550

Funders: Korea Research Institute of Bioscience and Biotechnology, Ministry of Science and ICT, South Korea