Restriction Landmark Genomic Scanning Analysis of Breast Cancer for Detection of Novel Amplicons
Annual rept. 15 Jul 96-14 JUl 97
MICHIGAN UNIV ANN ARBOR
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Genomic amplification is believed to be a pivotal event in the origin and progression of a variety of human tumors including breast cancers. Recently, a technical innovation potentially suitable for detecting candidate novel amplifications in tumors, called restriction landmark genomic scanning, has been described by Hyashizaki and co-workers. This is a two-dimensional DNA restriction analysis technique by which a highly reproducible constellation of -2,000 spots each corresponding to a specific genetic locus can be surveyed. As spot intensity is proportional to gene copy number in this system, amplified loci in tumor-derived DNAs are readily detected. Using this approach, several investigators including ourselves have demonstrated both previously identified and novel amplicons in several tumor types. A derivative technology allows direct cloning of amplified sequences from DNA isolated from excised spots. I propose here to study up to 20 breast tumor specimens in this way. Candidate novel amplicons identified will be cloned initial characterization of these amplicons will entail their genomic localization using fluorescence in situ hybridization.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research