Integration procedure localized multiple copies of a gene of interest in a strain of Yarrowia

Integration procedure localized at least three copies of a gene of interest into the genome of a strain of Yarrowia comprising the steps of: a) placing in culture of a strain of Yarrowia comprising at least three deletions selected from Ura3 -302, Leu2-270, Gut2-744 and Ade2-844 corresponding phenotype associated with each of these deletions an auxotrophy for said strain, independently of each other of these at least three genes; b) transforming said strain of Yarrowia auxotroph with at least three recombinant vectors comprise selectable markers that allow for said strain of Yarrowia, complementation of auxotrophy resulting from each of said deletions, said recombinant vectors each comprising : i) the sequence of said gene of interest, ii) a selection marker, and iii) two DNA sequences which frame the sequence of said gene of interest and said selectable marker, the said DNA sequences homologous to the sequences corresponding to the ends of the integration site located in the genome of said strain of Yarrowia so that integration localized said recombinant vector by homologous recombination is allowed; selection on minimal medium, yeasts having integrated the said at least three recombinant vectors.