NOVEL METHOD FOR INTEGRATING GENES AT SPECIFIC SITES IN MAMMALIAN CELLS VIA HOMOLOGOUS RECOMBINATION AND VECTORS FOR ACCOMPLISHING THE SAME

摘要

A METHOD FOR ACHIEVING SITE SPECIFIC INTEGRATION OF A DESIRED DNA AT A TARGET SITE IN A MAMMALIAN CELL VIA HOMOLOGOUS RECOMBINATION IS DESCRIBED. THIS METHOD PROVIDES FOR THE REPRODUCIBLE SELECTION OF CELL LINES WHEREIN A DESIRED DNA IS INTEGRATED AT A PREDETERMINED TRANSCRIPTIONALLY ACTIVE SITE PREVIOUSLY MARKED WITH A MARKER PLASMID. THE METHOD IS PARTICULARLY SUITABLE FOR THE PRODUCTION OF MAMMALIAN CELL LINES WHICH SECRETE MAMMALIAN PROTEINS AT HIGH LEVELS, IN PARTICULAR IMMUNO-GLOBULINS. NOVEL VECTORS AND VECTOR COMBINATIONS FOR USE IN THE SUBJECT CLONING METHOD ARE ALSO PROVIDED.(FIG. 3)