Abstract

ROSAS-QUIJANO, Raymundo  and  GAILLARDIN, Claude. The system Cre/loxP as a tool in the molecular study of Yarrowia lipolytica. Rev. Mex. Mic [online]. 2011, vol.33, pp.17-27. ISSN 0187-3180.

The non-conventional yeast Yarrowia lipolytica has been broadly studied, its whole genome sequence is known and public, and there are many molecular tools for study itself. However, their genetic markers are limited. Therefore, we need to recycle these markers. Taking advantage of Y. lipolytica has the ability for express different sources of recombinant protein, we constructed one replicating plasmid for Cre1 recombinase from one replicative plasmid under the direction of a strong promoter. The recombinase activity was evaluated in mutants obtained by transposition. The mTnYl1plasmid includes two Lox at ends who are target sequences of phage P1 recombinase. Our results indicated high efficiency and specificity of the recombinase activity. Based on this strategy we replaced the URA3 selective marker gene contained in mTnYl1 and then we sequenced the segments generated after recombinase activity to corroborate its proper action. This strategy is simple as well as a powerful molecular biology tool for Y. lipolytica study under selection and use of one single marker.

Keywords : heterologous expression; recombinase; genetic marker.

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