Transformation of mammalian cells with an amplifiable dominant-acting gene.

Abstract

A mutant hamster gene coding for an altered dihydrofolate reductase was transferred to wild-type cultured mouse cells by using total genomic DNA from methotrexate-resistant Chinese hamster ovary A29 cells as donor. By demonstrating the presence of hamster gene sequences in transformants, direct evidence for gene transfer was provided. Transformants selected for increased resistance to methotrexate contain increased amounts of the newly transferred gene. This mutant dhfr gene was used to introduce the Escherichia coli antibiotic resistance plasmid pBR322 into animal cells. Amplification of the dhfr sequences results in amplification of the pBR322 sequences as well. The use of this gene may allow the introduction and amplification of virtually any genetic element in various new cellular environments.