Stable, high-level expression of a carcinoembryonic antigen-encoding cDNA after transfection and amplification with...

The introduction and expression of cloned genes in a wide variety of animal cells requires the convenient use of dominant selectable markers. Very few of these markers can be amplified in copy number, a necessary feature if variable and high-level expression of the gene of interest is required. We describe the successful dominant transfection and amplification of a vector containing the Escherichia coli asparagine synthetase (AS)-encoding gene, asnA, transfected into a variety of human and rodent cell lines. An unlinked co-transfected expression vector containing the CEA cDNA, encoding human carcinoembryonic antigen, can be co-amplified with the asnA marker leading to extremely high levels of CEA synthesis. In addition, we show that the expression of both the asnA marker and the co-transfected CEA construct are stable in normal and amplified transfectants after prolonged culture in the absence of selective pressure.