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. 2019 May 31;12(6):1249–1259. doi: 10.1111/1751-7915.13427

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© 2019 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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Synthetic pathway assembly using the Golden Gate toolkit and the resulting expression in Y. lipolytica. A. Schematic representation of a three‐gene assembly, composed of Y. lipolytica xylulokinase (ylXK), Y. lipolytica xylitol dehydrogenase (ylXDH) and Y. lipolytica xylose reductase (ylXR) genes, that allows Y. lipolytica to grow on xylose. B. Y. lipolytica clones after transformation with the xylose cassette that were grown in a plate containing only xylose as the carbon source; clones were grown in a plate containing glucose as the carbon source for the control. The xylose cassette was correctly expressed in 79% of JMY1212 transformants. WT: wild type. Xyl: xylose. Glc: glucose. Ctrol+: positive control, which was the Y. lipolytica strain expressing the three genes developed via the standard cloning method and whose xylose utilization was verified (Ledesma‐Amaro et al., 2016).