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Identification of BRCAl and 2 Other Tumor Suppressor Genes on Chromosome 17 Through Positional Cloning

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Annual rept 1 Jul 96-30 Jun 97

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During the past year our project has evolved dramatically and significant progress has been make with respect to our overall goals of defining the genetic changes that distinguish normal cells from tumor cells of the breast. The most important evolution of the project is our change in strategy from identifying presumed tumor suppressor genes defined by loss of heterozygosity LOH using a positional cloning approach, to ascertaining and comparing genetic expression profiles from surgical specimens or cells grown in tissue culture using hybridization of cDNA from our tissues to microarrays of interesting genes. This is a very new technology and we are using a novel micro arraying and scanning instrument recently acquired by our laboratory in collaboration with Molecular DynamicsAmersham. Preliminary experiments suggest that the sensitivity of our new microarray system is sufficiently high to allow us to detect variance in gene expression levels with high precision even under complex hybridization conditions. We have also applied the Differential Display protocol to identify three genes with altered expression in a model system of normal and neoplastic epithelial colonocytes. As an alternative to determining biologic effects of specific genes, we have tested and selected an amphoteric retroviral conditional expression system, which replaces previous experiments using antisense oligos. We are confident that the modernization of our research project will facilitate the identification of genes critical to the development and progression of breast cancer.

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  • Biochemistry
  • Genetic Engineering and Molecular Biology
  • Anatomy and Physiology
  • Medicine and Medical Research

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