GENERATION OF PRODUCTION STRAINS THAT EFFICIENTLY EXPRESS NUCLEAR TRANSGENES

摘要

The present invention relates to a method of generating eukaryotic cells suitable for the expression of transgenes in said cells comprising (a) introducing a nucleic acid encoding a selectable marker responsive to a selecting agent into the nucleus of cells, wherein the level of expression of said selectable marker is proportional to the level of phenotypic responsiveness to said selecting agent; (b) selecting, among the cells obtained in step (a), for cells with a detectable expression of said selectable marker; (c) optionally propagating the cells selected for in step (b); (d) mutagenizing the cells selected for in step (b) or propagated in step (c) or allowing for the appearance of spontaneous mutations in the cells selected for in step (b) or propagated in step (c); and (e) selecting for cells displaying an increased expression of said selectable marker compared to the expression obtained in step (b). The present invention furthermore relates to a eukaryotic cell produced by the method of the present invention, a method of producing a compound of interest in a cell produced with the method of the present invention comprising (a) introducing a nucleic acid encoding (i) the compound of interest which is a protein or an RNA; or (ii) a protein necessary to synthesize said compound of interest; and optionally a selectable marker responsive to a selecting agent into said cell; (b) expressing said protein in the cell; and (c) isolating the compound of interest produced; and a kit comprising (a) a cell obtainable by the method of the invention and optionally a vector optimized for protein expression in said cell; or (b) the cell of the invention.