Use of a Transfected and Amplified Drosophila Heat Shock Promoter Construction for Inducible Production of Toxic Mouse C-Myc Proteins in CHO Cells

摘要

After transfection and selection with methotrexate, CHO cell lines were established which contained up to 2000 copies of an expression vector for c-myc protein. The vector contained the Drosophila heat shock protein 70 (hsp70) promoter fused with the coding region of the mouse c-myc gene. Incubation of cells for up to 3 hours at 43 sup 0 C resulted in at least a 100-fold induction of recombinant c-myc mRNA. When cells were shifted back to 37 sup 0 C, within 1 to 4 hours, this RNA was translated into protein to yield about 250 mu g per 10 sup 9 cells. Cells died a few hours later, suggesting that high concentrations of intracellular c-myc are cytotoxic. 47 refs., 5 figs. (ERA citation 12:050261)