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A Functional Screen for Germ Line p53 Mutations Based on Transcriptional Activation¹

Division of Molecular Genetics, Massachusetts General Hospital Cancer Center, Charlestown, Massachusetts 02129 [T. F., N. B., J. K., Y-S. N., P. R., S. H. F.], and Department of Pediatrics, Division of Hematology-Oncology, The Children's Hospital, Dana-Farber Institute, Harvard Medical School, Boston Massachusetts, 02114 [S. H. F.]

² To whom requests for reprints should be addressed.

Germ line p53 mutations represent a genetic predisposition for cancer development. At the present time, their detection requires extensive work and their functional significance must be documented. Therefore, we have designed a simple biological assay which detects functionally significant germ line p53 mutations. This assay is based on the cloning of the patient's p53 complementary DNA into a eukaryotic expression vector followed by the cotransfection into human cells of the recombinant vector with a reporter plasmid for the transcriptional activity of p53. This assay potentially offers a powerful method to screen fibroblasts or lymphocytes from patients for germ line mutations which inactivate the p53 tumor suppressor gene.

¹ This work was supported in part by John T. Merk Fund, the Lucille P. Markey Foundation, and the Robert Steel Foundation.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Received 10/ 7/92. Accepted 11/ 6/92.

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