Patent ID: 6524824
Filing Date: 2003-02-25
Classification: A61K,C07K,C12N

Abstract:
A method for detecting a eukaryotic cell expressing an endogenous gene or portion thereof encoding an integral membrane protein or portion thereof, said method comprising (a) introducing into said cell in vitro a vector comprising in operable combination: a promoter operably linked to an exon sequence comprising a start codon, a signal sequence required for secretion of said protein, and an epitope tag followed by an unpaired splice donor site; wherein said vector is non-homologously incorporated into the genome of said cell and the unpaired splice donor site on the vector is spliced to a splice acceptor site in the endogenous gene encoding said integral membrane protein, wherein said splice acceptor site is 5â€² to one or more exons that encode said integral membrane protein or portion thereof, wherein a fusion protein is produced comprising the vector-encoded epitope tag linked to said integral membrane protein or portion thereof, wherein said fusion protein is embedded in the cell membrane such that the epitope tag is detectable on the cell surface, and wherein said promoter is not identical to the promoter that is naturally operably-linked to said endogenous gene; and (b) screening said cells for said epitope tag.