abstract |
A heterologous gene coding sequence is inserted into a target endogenous gene in a eukaryotic cellular host cell genome and expressed, by transforming the host cell with a vector comprising a DNA construct comprising the sequence: 5' X-A-P-B-Q-C-Y 3' in which X and Y are substantially homologous with separate sequences from the target endogenous gene and are of sufficient length to undergo homologous recombination with the host cell genome so as to insert the A-P-B-Q-C sequence into the host cell genome; P is an internal ribosome entry site (IRES); Q is the heterologous gene coding sequence; and A, B and C are, separately, additional DNA sequences optionally present in the construct. Also described are cells, non-human animals, descendants and vectors comprising the heterologous coding sequence, and use of the construct in manufacture of a vector. |