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首页> 外文会议>Conference on optical biopsies and microscopic techniques >Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH
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Localization of specific sequences and DNA single-strand breaks in individual UV-A-irradiated human lymphocytes by COMET FISH

机译：特定序列的定位和DNA单链由彗星鱼中的单独的UV-A辐照人淋巴细胞中断

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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-, $alpha@- satellite-, telomere-, whole chromosome-, single copy- and region specific DNA probes have been used to investigate whether the UV-A induced DNA single strand breaks are distributed randomly all over the human genome or induced at specific sites ('hot spots'). In the investigated human peripheral blood lymphocytes all but one centromere reveal low sensitivity for UV-A irradiation (500 kJ/m$+2$/), while telomeres are randomly distributed over COMET heads and tails. The human chromosome 1 is fractionated by irradiation, but remains in the COMET head, indicating an only moderate degree of fractionation. Among three tested single copy probes, c- myc, p53 and p58, the p53 gene located on chromosome 17p13.1 and the p58 gene (1p36) appear to be located in UV-A stable regions of the human genome in 95% of 65 investigated lymphocytes. In contrast, the c-myc proto-oncogene (8q24) is found in the COMET tail in 90% of the 27 investigated lymphocytes and thus appears to be more sensitive to UV-A irradiation.
机译：彗星测定，一种单细胞电泳技术，其允许根据尺寸分离电泳分级的DNA的单细胞电泳技术已与原位杂交（鱼类）的荧光组合，其允许定位特定基因或基因区域。这种组合（彗星鱼）允许在单细胞水平处检测人淋巴细胞基因组的特定区域中的DNA单链断裂。各种类型的DNA探针，例如， Centromere-，$ alpha @ - 卫星 - ，端粒，整个染色体 - ，单拷贝和区域特异性DNA探针用于研究UV-A诱导的DNA单链断裂是否随机分布在人类基因组或诱导在特定网站（'热点'）。在调查的人外周血淋巴细胞中，除了一个Centromere揭示了UV-A辐射的低灵敏度（500 KJ / m + 2美元/），而端粒则会随机分布在彗星头和尾部。人染色体1通过辐射分馏，但保留在彗星头中，表明仅适中的分馏。在三种测试的单拷贝探针中，C-myc，p53和p58，位于染色体17p13.1和p58基因（1p36）上的p53基因似乎位于UV-A稳定的人类基因组中，在65％的95％中调查淋巴细胞。相反，在27所研究的淋巴细胞的90％的彗星尾部发现C-Myc原癌基因（8Q24），因此对UV-A辐射似乎更敏感。

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