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Production of Recombinant Rat Viruses as a Method of Oncogene Isolation in Coculture Medium

Department of Biochemical Genetics, Medical Research Institute, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, Tokyo 113 [S-i. K.]; Division of Cell Biology, Institute for Comprehensive Medical Science, Fujita-Gakuen Health University, Toyoake-shi, Aichi 470-11 [N. K.]; Molecular Oncology Laboratory, Chemical and Physical Research Institute, Hirosawa, Wako-shi, Saitama 351 [Y. I.]; and Mitsubishi-Kasei Institute of Life Sciences, 11 Minamiooya, Machida-shi, Tokyo 194 [S-i. K.], Japan

A simple oncogene isolation was proved using the SD1-T rat embryonic cell line. The SD1-T cell line, which releases endogenous rat leukemia virus, was cocultured with (a) normal rat kidney cells transformed by cloned v-mos DNA, (b) the rat mammary tumor cell line (63SP), or (c) normal rat kidney cells transformed by 63SP DNA. Within 1 mo, oncogenic viruses were recovered from all three coculture supernatants. During this period, increase of oncogenic transcripts was observed in the cocultured cells. The oncogenic viruses appeared to contain the mos gene in cocultures (a) and ras-related sequences in cocultures (b) and (c). The emergence of virus containing mos from mos DNA-mediated normal rat kidney transformants demonstrated "rescue" of the active cellular oncogene by the rat leukemia virus. This coculture system seems to facilitate "rescue" of oncogenes functioning in the tumor and transformed cells.

¹ To whom requests for reprints should be addressed, at Mitsubishi-Kasei Institute of Life Sciences, 11 Minamiooya, Machida-shi, Tokyo 194, Japan.

Received 4/24/87. Revised 8/ 5/87. Accepted 8/14/87.

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