| 核分离-滤膜洗脱技术同时检测植物DNASSB与DSB |
| 摘要点击 1973 全文点击 2287 投稿时间：2000-01-14 |
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| 中文关键词 DNA链断裂 核分离-微孔滤膜洗脱法 60Co-γ 辐射 大麦 生物标志物 |
| 英文关键词 DNA breaks nuclei isolation filter elution assay 60Co-γ radiation barley biomarker |
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| 中文摘要 |
| 采用核分离 -滤膜洗脱技术 ,将分离的细胞核在微孔滤膜上裂解后相继用中性溶液和碱性溶液洗脱 ,研究了60Co- γ辐射引发的大麦种胚细胞 DNA断裂 .结果显示 ,50 Gy的 60Co- γ辐射主要引发 DNA单链断裂 (DNA SSB) ;50～100Gy的辐射引发的DNA断裂包括单链断裂 (SSB)与双链断裂 (DSB) ;当辐射剂量大于 100Gy时 ,DNA断裂的增加部分则以 DSB为主 .所用的实验方法适用于质地致密的植物材料并可在一次实验中同时检测 DNA的 SSB与 DSB,为植物细胞的 DNA链断裂作为生物标志物应用于环境诱变因子监测提供了基础 . |
| 英文摘要 |
| Barley embryo DNA breaks in imbibing intact barley seeds induced by 60Co-γ irradiation were studied by a nuclei isolation filter elution assay,which permits the simultaneous detection of DNA double strand breaks(DSB)and single strand breaks(SSB)of compact plant donor.This assay based on the procedures of 4 h of neutral solution elution followed by 4 h of alkaline solution elution after lysing the free nuclei instead of intact cells by proteinase K on the 0.45μm pore size filters.This approach offers a new kind of biomarker of DNA breakage in plant cells in the monitoring of mutagenic agent in the environment.The results indicated that when the barley seeds were exposed to different dosages of γ-ray,DNA breaks in embryo cells were induced in the following manner:(1)production mainly of SSB by 50Gy;(2)production of both SSB and DSB between 50 and 100Gy;and (3)production of DNA breaks with the increment in DSB by the dosage over 100Gy. |
