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. 2022:2513:205-220.
doi: 10.1007/978-1-0716-2399-2_12.

Golden Gate Multigene Assembly Method for Yarrowia lipolytica

Macarena Larroude  1 , Jean-Marc Nicaud  1 , Tristan Rossignol  2
Affiliations

Golden Gate Multigene Assembly Method for Yarrowia lipolytica

Macarena Larroude et al. Methods Mol Biol. 2022.

Abstract

The oleaginous yeast Yarrowia lipolytica has emerged as a powerful alternative for biolipid production due to its high capacity for lipid accumulation. Genetic engineering and synthetic biology are promoted forward to improve production and reroute metabolism for high-value compound synthesis. In this context, efficient, modular, and high-throughput compatible cloning and expression system are required to speed up and rationalize research in this field. Here, we present the fast and modular Golden Gate cloning strategy for the construction of multigene expression vectors and their transformation into Y. lipolytica. As an example, we used the heterologous expression of the carotenoid pathway by cloning three genes involved in this pathway in only one vector allowing reaching production of β-carotene after a single transformation.

Keywords: Gene assembly; Gene expression; Golden Gate; Synthetic biology; Yarrowia lipolytica; β-carotene production.

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