Method for the targeted integration of multiple copies of a gene of interest in a yarrowia strain
US8748129B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Feb 5, 2009 |
| Grant date | Jun 10, 2014 |
| Priority date | — |
| Expiry date | Sep 20, 2029 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N15/815
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
This invention concerns a method for the targeted integration of at least three copies of a gene of interest in the genome of a Yarrowia strain including the steps of: (a) cultivating a Yarrowia strain, said strain including a deletion among at least three genes, the phenotype associated with each of these deletions corresponding to an auxotrophy or to a dominant character for this strain; (b) transforming said Yarrowia strain thus obtained with at least three recombinant vectors that include selection markers allowing, for this strain, the complementation of auxotrophy and, potentially, of the dominant character resulting from each of these deletions; and (c) selecting, on a minimum medium, the yeasts having integrated said at least three recombinant vectors. This invention also includes a method for producing a polypeptide of interest using this method as well as a method for obtaining a modified Yarrowia strain including a deletion among at least three genes, the phenotype associated with each of these deletions corresponding to an auxotrophy or to a dominant character for this strain.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.