No document available.
Abstract :
[en] Selectable markers are a central component of genome edition technologies. In the yeast Yarrowia lipolytica, these markers are traditionally based on antibiotic resistance (hygromycin B) or auxotrophy (e.g., leucine, uracil). However, the use of the former is impaired by a high level of spontaneous resistance, and the use of the latter by continuous complementation of the culture medium or restoration of prototrophy to the strains. As an alternative, genes related to the catabolism of carbon sources, or “catabolic selectable markers”, present the advantage of not being involved in essential metabolic pathways. The recently identified EYK1, encoding erythrulose kinase, can serve as an efficient catabolic selectable marker for genome editing in Y. lipolytica. Compared to auxotrophic markers such as URA3 and LEU2, EYK1 increases the growth rate of transformants on selective medium and the efficiency of genome edition. The utility of the marker EYK1 in a replicative vector was also demonstrated. Besides, the cloning-free strategy developed here simplifies the construction of disruption cassettes for genome editing in Y. lipolytica.