An alternative assay system for the detection of transforming genes |
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| Authors: | Brown Robert; Brady Ged; Mattern Jurgen; Schutz Gunther |
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| Institution: | Institut für Zell- and Tumorbiologie, Deutsches Krebsforschungszentrum Im Neuenheimer Feld 280, 6900 Heidelberg, FRG
2Institut fur Experimentelle Pathologic, Deutsches Krebsforschungszentrum Im Neuenheimer Feld 280, 6900 Heidelberg, FRG |
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| Abstract: | We have developed an alternative assay system based on DNA mediatedgene transfer (DMGT) for the detection of functional transforminggenes which we find more sensitive and reliable than other assaysystems. This approach involves selecting in tissue culturefor cells which are expressing a drug-resistance marker co-transferredwith the tumour DNA thus isolating the small number of cellswhich have taken up and are expressing exogenously added DNA.The ability of transformed cells present in this drug-resistantcell population to form tumours in athymic (nu/nu) nude micewas then used to assay for transforming genes. Using NIH3T3and Rat 2 cells as recipients, tumours produced after DMGT withDNA from non-tumorgenic cells were only very rarely observed.Tumour induction was observed using DNA from a biopsy of a humanovary carcinoma and a spontaneous murine lung carcinoma. Twohuman retinoblastoma cell lines were negative for tumour induction.The transforming gene from the ovarian carcinoma was identifiedas c-K-ras2. |
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