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生态学杂志 ›› 2005, Vol. ›› Issue (8): 975-979.

• 技术与方法 • 上一篇    下一篇

单细胞凝胶电泳技术及在土壤生态毒理学中的应用

林爱军, 耿春女, 朱永官, 童依平   

  1. 中国科学院生态环境研究中心土壤环境研究室, 北京, 100085
  • 收稿日期:2004-04-19 修回日期:2004-06-23 出版日期:2005-08-10
  • 通讯作者: 朱永官
  • 基金资助:
    中国科学院知识创新重大项目(KZCX1-SW-19);国家重点基础研究发展规划项目(2002CB410808);中国科学院“百人计划”资助项目.

Single cell gel electrophoresis technique and its applications in soil ecotoxicology

LIN Aijun, GENG Chunn, ZHU Yongguan, TONG Yiping   

  1. Department of Soil Environmental Sciences, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China
  • Received:2004-04-19 Revised:2004-06-23 Online:2005-08-10

摘要: 单细胞凝胶电泳技术又称为彗星实验,是最近几年发展起来的一种快速、简单、灵敏、可靠的检测细胞核DNA损伤的技术。总结了近几年来单细胞凝胶电泳技术的发展、原理、方法及其应用,并指出其下一步的发展趋势。彗星实验中,镶嵌于琼脂糖中的细胞核在电场中向正极移动,因细胞核与DNA片段迁移速率不同,而形成类似“彗星”的图像。目前采用的彗星实验有多种,可以检测诸如DNA双链断裂、单链断裂、碱不稳定位点等多种类型的DNA损伤。碱性彗星实验因其高灵敏度而被广泛采用。彗星实验的主要步骤包括细胞核悬浮液的获得、彗星电泳胶板制备、细胞裂解、DNA变性解旋、电泳、中和、染色和观察等。目前彗星实验广泛应用于各个研究领域,近年来开始用于环境污染的基因毒性研究和生物监测,并取得了迅速发展。

关键词: 日光温室, 地面覆盖, 嫁接黄瓜, 产量, 土壤环境

Abstract: The single cell gel electrophoresis technique (comet assay) is a simple,sensitive,reliable,and rapid method to detect DNA damage in eukaryotic cells.The paper reviewed the principles,advances and methodology of comet assay,and its applications in soil ecotoxicology,with its development in future predicted.Nuclei embedded in low-melting-point agrose in an agar gel sandwich migrated towards the anode under an electrical current with a different rate from that of DNA strands presented the appearance of a "comet".Different techniques are available with different sensitivity to detect DNA damages such as double strand breaks,single strand breaks,delay DNA repair sites and alkali labile sites.The basic procedure of the technique included cell suspension preparation,slide preparation,lysis,unwinding,electrophoresis,neutralization,dying and view.The method has been widely used in several areas in the past few years,especially in genotoxicology research in environmental biomonitoring.

Key words: Solar greenhouse, Soil surface mulching, Grafted cucumber, Yield, Soil environment

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