Abstract
Budding yeast, as a eukaryotic model organism, has well-defined genetic information and a highly efficient recombination system, making it a good host to produce exogenous chemicals. Since most metabolic pathways require multiple genes to function in coordination, it is usually laborious and time-consuming to construct a working pathway. To facilitate the construction and optimization of multicomponent exogenous pathways in yeast, we recently developed a method called YeastFab Assembly, which includes three steps: (1) make standard and reusable genetic parts, (2) construct transcription units from characterized parts, and (3) assemble a complete pathway. Here we describe a detailed protocol of this method.
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Acknowledgments
This work was supported by the National Key Research and Development Program of China (2018YFA0900100), the National Natural Science Foundation of China (31725002, 31800069, and 31800082), Shenzhen Key Laboratory of Synthetic Genomics (ZDSYS201802061806209), Shenzhen Science and Technology Program (KQTD20180413181837372), Guangdong Provincial Key Laboratory of Synthetic Genomics (2019B030301006), and Bureau of International Cooperation, Chinese Academy of Sciences (172644KYSB20180022, 172644KYSB20170042).
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Jiang, S., Luo, Z., Dai, J. (2021). Use YeastFab to Construct Genetic Parts and Multicomponent Pathways for Metabolic Engineering. In: Xiao, W. (eds) Yeast Protocols. Methods in Molecular Biology, vol 2196. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0868-5_13
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DOI: https://doi.org/10.1007/978-1-0716-0868-5_13
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0868-5
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