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MONITORING ULTRAVIOLET‐B‐INDUCED DNA DAMAGE IN INDIVIDUAL DIATOM CELLS BY IMMUNOFLUORESCENT THYMINE DIMER DETECTION 1
Author(s) -
Buma Anita G. J.,
Hannen Erik J.,
Roza Len,
Veldhuis Marcel J. W.,
Gieskes Winfried W. C.
Publication year - 1995
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.0022-3646.1995.00314.x
Subject(s) - pyrimidine dimer , biology , dna , thymine , dna damage , flow cytometry , fluorescence , microbiology and biotechnology , staining , nucleolus , biophysics , fluorescein isothiocyanate , in situ , nuclear dna , fluorescence microscope , biochemistry , cytoplasm , chemistry , mitochondrial dna , genetics , gene , optics , physics , organic chemistry
We developed a method to investigate the effect of ultraviolet‐B radiation (UVBR) on the formation of thy‐mine dimers in microalgal DNA that can be used for both laboratory and in situ research. Antibody labeling of dimers was followed by a secondary antibody (fluorescein isothiocyanate) staining to allow visualization of DNA damage with flow cytometry or fluorescence microscopy. Thymine dimer‐specific fluorescence in nuclear DNA of the marine diatom Cyclotella sp. was linearly related to the UVBR dose. Simultaneous measurements of cellular DNA content showed that the vulnerability of G2 cells to DNA damage did not differ significantly from the vulnerability of G1 cells. The formation and removal of thymine dimers in Cyclotella sp. cells was monitored for 3 consecutive days at two realistic UVBR irradiance levels. Thy‐mine dimers were removed within 24 h when exposed to a saturating photosynthetically active radiation intensity following the UVBR treatment. This new method allows the study of UVBR‐induced DNA damage on a cell‐to‐cell basis. It is also feasible for field studies because cells remain intact and can be recognized readily after antibody treatment .