Construction of a retroviral cDNA version of the int-1 mammary oncogene and its expression in vitro.
The int-1 mammary oncogene is activated by proviruses of the Mouse Mammary Tumor Virus in many different mammary tumors. We have inserted a genomic fragment containing the protein-encoding domain of the gene into the retroviral shuttle vector pZIPneoSV(X)1. After one round of virus replication we recovered recombinant proviral DNA containing a correctly spliced copy of int-1. In vitro transcription of this cDNA version of int-1 using SP6 polymerase and translation in a reticulocyte lysate yielded a protein of approximately 37,000 daltons. High expression of int-1 in NIH-3T3 cells infected with recombinant virus did not lead to morphological transformation.
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004247:DNA / Q000378:metabolism*; D004262:DNA Restriction Enzymes; D005814:Genes, Viral; D007668:Kidney; D008325:Mammary Neoplasms, Experimental / Q000382:microbiology; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D051379:Mice; D008815:Mice, Inbred Strains; D009857:Oncogenes; D014158:Transcription, Genetic; D014162:Transfection
D004247:DNA; D004262:DNA Restriction Enzymes