Detection of Mutations Using a Novel Endonuclease
Annual rept. 1 Jul 1997-30 Jun 1998,
FOX CHASE CANCER CENTER PHILADELPHIA PA
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The detection of mutations in the genes of breast cancer patients and their families is an important part of the effort to counter breast cancer. This laboratory has discovered a family of nucleases that has the ability to cut DNA at the sites of mismatches, such as those that are formed when a DNA containing a mutation is paired with a copy of the normal DNA. The enzyme we use is CEL I from celery. The CEL I mismatch detection CMD assay has shown high efficacy. In the work supported by this grant, we have tested the detection of mutations in the BRCAl gene of many research participants. CMD detected mutations using either the PCR products amplified from DNA, or the RT-PCR products amplified from their mRNA. The results showed that CMD will be useful in the genetic screening against breast cancer. Through this work, we obtained insights on approaches that will further enhance the sensitivity of CMD. These methods will be tested in the following period of research support by this grant.
- Genetic Engineering and Molecular Biology
- Medicine and Medical Research