Transformation of mammalian cells with an amplifiable dominant-acting gene.
We have transferred a mutant hamster gene coding for an altered dihydrofolate reductase 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 we have provided direct evidence for gene transfer. Transformants selected for increased resistance to methotrexate contain increased amounts of the newly transferred gene. We have used this mutant dhfr gene 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.
D000818:Animals; D002460:Cell Line; D002999:Clone Cells / Q000201:enzymology; D006224:Cricetinae / Q000235:genetics*; D003412:Cricetulus / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005347:Fibroblasts / Q000378:metabolism; D005799:Genes, Dominant; D005822:Genetic Vectors; D051379:Mice / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics*; D014170:Transformation, Genetic
D013762:Tetrahydrofolate Dehydrogenase