Selection of Transfected Mammalian Cells

Magnetic selection of transiently transfected cells.

An efficient and rapid method for selecting transiently transfected cells is described. A plasmid encoding for a neural cell-specific surface marker is co-transfected into mammalian cells along with the gene of interest. After uptake and expression of these two plasmids, the transfected cells are immuno-adsorbed to magnetic beads pre-coated with antibodies against the surface marker. These immu...

Effects of methotrexate on transfected DNA stability in mammalian cells.

The chromosomal locations, amounts, and level of expression of transfected, amplified c-myc and dihydrofolate reductase sequences were measured in cells cultured in the presence and absence of methotrexate. These studies show that the location and amount of transfected sequences, as well as the level of expression, were more variable when the cells were cultured in methotrexate.

High mutation frequency in DNA transfected into mammalian cells.

The lacI gene of Escherichia coli was used to score mutation in mammalian cells of simian virus 40-based recombinant DNA vectors that provide for replication and selection in both bacterial and mammalian cells. Plasmid DNA was introduced into COS7 simian cells by DEAE-dextran transfection, allowed to replicate in the mammalian cells, and then returned to E. coli for analysis. Mutants in lacI we...

Simultaneous selection and cloning of transfected eukaryotic cells.

Transcriptional repression by Drosophila and mammalian Polycomb group proteins in transfected mammalian cells.

The Polycomb group (Pc-G) genes are essential for maintaining the proper spatially restricted expression pattern of the homeotic loci during Drosophila development. The Pc-G proteins appear to function at target loci to maintain a state of transcriptional repression. The murine oncogene bmi-1 has significant homology to the Pc-G gene Posterior sex combs (Psc) and a highly related gene, Suppress...