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Clinical Chemistry 41: 1841-1843, 1995;
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Clinical Chemistry, Vol 41, 1841-1843, Copyright © 1995 by American Association for Clinical Chemistry

Detection of point mutations in T lymphocytes

AA van Zeeland, JG Jansen, GR Mohn, H Vrieling, AD Tates and PH Lohman
Department of Radiation Genetics and Chemical Mutagenesis, Leiden University, The Netherlands.

We exposed experimental animals to a series of alkylating agents that induced mutations at the X-linked hprt gene of T lymphocytes. We then isolated the mutant cells and analyzed the molecular nature of the mutations by amplification of hprt cDNA sequences with the use of reverse transcriptase PCR followed by DNA sequence analysis, and then correlated the mutational spectra obtained to the spectra of DNA adducts caused by the alkylating agents used. The nature of the base- pair changes causing the mutations was characteristic for the reaction pattern of the genotoxic agent with DNA. However, we also found a clear influence of DNA repair processes; i.e., in those cells that were able to remove certain types of DNA damage, the class of mutations expected from that type of damage was reduced.




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