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Proceedings Paper

Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH
Author(s): Claudia Bock; Alexander Rapp; Heike Dittmar; Shamci Monajembashi; Karl-Otto Greulich
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Paper Abstract

The COMET assay, a single cell electrophoresis technique which allows to separate electrophoretically fractionated DNA according to size has been combined with fluorescence in situ hybridization (FISH) which allows to localize specific genes or gene regions. This combination (COMET FISH) allows the detection of DNA single strand breaks in specific regions of the genome of human lymphocytes at the single cell level. Various types of DNA probes, e.g. centromere-, (alpha) - satellite-, telomere-, whole chromosome-, single copy- and region specific DNA probes have been used to investigate whether the UV-A induced DNA single strand breaks are distributed randomly all over the human genome or induced at specific sites ('hot spots'). In the investigated human peripheral blood lymphocytes all but one centromere reveal low sensitivity for UV-A irradiation (500 kJ/m2), while telomeres are randomly distributed over COMET heads and tails. The human chromosome 1 is fractionated by irradiation, but remains in the COMET head, indicating an only moderate degree of fractionation. Among three tested single copy probes, c- myc, p53 and p58, the p53 gene located on chromosome 17p13.1 and the p58 gene (1p36) appear to be located in UV-A stable regions of the human genome in 95% of 65 investigated lymphocytes. In contrast, the c-myc proto-oncogene (8q24) is found in the COMET tail in 90% of the 27 investigated lymphocytes and thus appears to be more sensitive to UV-A irradiation.

Paper Details

Date Published: 19 January 1999
PDF: 11 pages
Proc. SPIE 3568, Optical Biopsies and Microscopic Techniques III, (19 January 1999); doi: 10.1117/12.336836
Show Author Affiliations
Claudia Bock, Institut fuer Molekulare Biotechnologie e.V. (Germany)
Alexander Rapp, Institut fuer Molekulare Biotechnologie e.V. (Germany)
Heike Dittmar, Institut fuer Molekulare Biotechnologie e.V. (Germany)
Shamci Monajembashi, Institut fuer Molekulare Biotechnologie e.V. (Germany)
Karl-Otto Greulich, Institut fuer Molekulare Biotechnologie e.V. (Germany)

Published in SPIE Proceedings Vol. 3568:
Optical Biopsies and Microscopic Techniques III
Irving J. Bigio; Herbert Schneckenburger; Jan Slavik; Katarina Svanberg M.D.; Pierre M. Viallet, Editor(s)

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