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. 1986 Jan 24;14(2):693–702. doi: 10.1093/nar/14.2.693

Construction of a retroviral cDNA version of the int-1 mammary oncogene and its expression in vitro.

F A Rijsewijk, M van Lohuizen, A van Ooyen, R Nusse
PMCID: PMC339458  PMID: 3003692

Abstract

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.

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