Patent ID: 6740503
Filing Date: 2004-05-25
Classification: A61K,C07K,C12N

Abstract:
A method for expression of a protein encoded by a genomic DNA fragment, said method comprising:(A) transfecting a host cell in vitro with a vector comprising: (a) a promoter operably linked to: (i) an exon; and (ii) a splice donor that defines the 3â€² end of the exon; and (b) one or more selectable markers; and (c) a genomic DNA fragment, wherein said vector is not integrated into the genome of the host cell or, when integrated into the genome of the host cell, does not integrate by homologous recombination into or around the gene that is expressed on the genomic DNA fragment, wherein said promoter, exon and splice donor are operably linked to a gene or portion thereof in said genomic DNA fragment, said gene or portion thereof encoding a protein and comprising one or more splice acceptor sequences, wherein said exon and/or promoter is heterologous to said gene or portion thereof on the genomic DNA fragment, wherein said exon lacks a functional translation start codon and wherein said splice donor in a transcript containing said exon is spliced to said splice acceptor on the genomic DNA fragment, wherein a spliced transcript is produced comprising said exon and said gene or portion thereof encoding a protein, and wherein said transcript is translated in said cell.