Inducible overproduction of the mouse c-myc protein in mammalian cells
We have made Chinese hamster ovary (CHO) cell lines that contain up to 2000 copies of the coding region of the mouse c-myc gene fused to the promoter of the Drosophila gene (hsp70) encoding a Mr 70,000 heat shock protein. Incubation of these cells at 43 degrees C results in an estimated 100-fold induction of c-myc mRNA. Translation of this mRNA occurs when the cells are returned to 37 degrees C, and during the first 3 hr of recovery at 37 degrees C, the c-myc protein is one of the most abundantly synthesized proteins in the cells. The products of the induced c-myc gene are phosphoproteins of apparent Mr 64,000, 66,000, and 75,000. Induced cells die, suggesting that elevated levels of c-myc are cytotoxic. Amplification of genes placed under control of the Drosophila hsp70 promoter may provide a general method for inducibly over expressing proteins in mammalian cells.
- URL: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=3526329
Keywords: Animals ; Cell Survival ; Cloning ; Molecular ; Gene Amplification ; Gene Expression Regulation ; Genetic Engineering ; Heat-Shock Proteins/genetics ; Mice ; Promoter Regions (Genetics) ; Proto-Oncogene Proteins/*genetics ; RNA ; Messenger/metabolism ; Temperature
Record created on 2007-06-05, modified on 2016-08-08