Patent ID: 6528314
Filing Date: 2003-03-04
Classification: A01K,A61K,C07K,C12N

Abstract:
An in vitro process for providing a recombinant, heterologous gene in a genome of a mammalian cell, wherein the process comprises:(A) providing a mammalian cell having a recipient gene in its genome, wherein said recipient gene comprises complementing DNA comprising a first nucleotide sequence and a second nucleotide sequence downstream of said first nucleotide sequence; (B) transfecting said mammalian cell in vitro with a DNA comprising: (1) a third nucleotide sequence homologous to said first nucleotide sequence; (2) a fourth nucleotide sequence homologous to said second nucleotide sequence; and (3) a DNA sequence heterologous with respect to DNA in said recipient gene, wherein said heterologous DNA sequence is between said third and said fourth nucleotide sequences and said heterologous DNA sequence comprises a first insertion DNA sequence and a second insertion DNA sequence, wherein said first insertion DNA sequence comprises a first coding sequence that encodes a first product that does not confer resistance to a selection agent involved in the selection of transformants, and a regulatory sequence for regulating the expression of said first product, and said second insertion DNA sequence comprises a second coding sequence that encodes a second product that confers resistance to a selection agent involved in the selection of transformants, and a promoter allowing the expression of said second product in said mammalian cell; and (C) selecting a transfected cell in which said heterologous DNA sequence has been inserted between said first and said second nucleotide sequences in said recipient gene by homologous recombination with said third and said fourth nucleotide sequences to thereby provide a mammalian cell containing in its genome the recombinant, heterologous gene, and in addition, the second insertion DNA sequence encoding the selective agent.