AN AUTOCATALYTIC EXPRESSION SYSTEM FOR REGULATED PRODUCTION OF RECOMBINANT PROTEIN IN MAMMALIAN-CELLS

An autocatalytic inducible mammalian expression system was established . This system is composed of two sets of vectors: one carries a select able marker gene and the other carries a bicistronic expression unit c onsisting of a target gene, an internal ribosomal entry site, and a te tracycline-controlled transactivator gene. Both the selectable marker and the bicistronic unit are controlled by the tetracycline-responsive promoter (PhCMV-1). When the two vectors are cotransfected into host cells, only a single selection round in the absence of tetracycline i s necessary to generate clones expressing the target gene. The express ion level is high and easily regulated by tetracycline. Combination wi th a gene amplification system may allow further enhance the foreign g ene expression. Because the PhCMV-1 promoter is relatively independen t of cellular regulation signals, this system is expected to work in a wide variety of cell types. (C) 1997 Academic Press.