摘要 |
A method for selecting recombinant host cells expressing high levels of a desired protein is described. This method utilizes eukaryotic host cells harboring a DNA construct comprising a selectable gene (preferably an amplifiable gene) and a product gene provided 3' to the selectable gene. The selectable gene is positioned within an intron defined by a splice donor site and a splice acceptor site and the selectable gene and product gene are under the transcriptional control of a single transcriptional regulatory region. The splice donor site is generally an efficient splice donor site and thereby regulates expression of the product gene using the transcriptional regulatory region. The transfected cells are cultured so as to express the gene encoding the product in a selective medium comprising an amplifying agent for sufficient time to allow amplification to occur, whereupon either the desired product is recovered or cells having multiple copies of the product gene are identified.
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