Applied Microbiology and Biotechnology

, Volume 103, Issue 2, pp 963–971 | Cite as

Selectable marker recycling in the nonconventional yeast Xanthophyllomyces dendrorhous by transient expression of Cre on a genetically unstable vector

  • Ning Zhang
  • Jiaxin Li
  • Fuli Li
  • Shi’an WangEmail author
Methods and protocols


Selectable marker recycling is a basic technique in bioengineering. However, this technique is usually unavailable in non-model microorganisms. In this study, we proposed a simple and efficient method for selectable marker recycling in the astaxanthin-synthesizing yeast Xanthophyllomyces dendrorhous. This method was based on a Cre-loxP system, in which the transient expression of the Cre recombinase was controlled by a genetically unstable vector independent of episomal plasmids and inducible promoters. The selectable markers in single-gene locus and multigene loci were removed along with the loss of the Cre vector with a ratio of 100% and 29%, respectively. The significance of the method was highlighted by the finding that stable autotrophic mutants were not readily obtained in X. dendrorhous. Comparative studies in X. dendrorhous and the non-homologous end joining dominant yeast Yarrowia lipolytica suggested that the method could be universally used in homologous recombination dominant yeasts.


Cell factory Yeast Cre-loxP system Marker recycling Homologous recombination 



We thank the Natural Science Foundation of China and Shandong Provincial Natural Science Foundation for financial support.


This study was supported by the Natural Science Foundation of China (No. 31670054 and No. 21676159) and Shandong Provincial Natural Science Foundation (ZR2017ZB0209).

Compliance with ethical standards

Conflict of interests

The authors declare that they have no conflict of interests.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_9496_MOESM1_ESM.pdf (762 kb)
ESM 1 (PDF 761 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Shandong Provincial Key Laboratory of Synthetic Biology, Key Laboratory of Biofuels, Qingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoChina
  2. 2.University of Chinese Academy of SciencesBeijingChina

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