遗传 ›› 2008, Vol. 30 ›› Issue (5): 659-662.doi: 10.3724/SP.J.1005.2008.00659

• 技术与方法 • 上一篇    

检测DNA-蛋白质交联的新方法

苏来, 宋宏宇   

  1. 国家沈阳新药安全评价中心, 沈阳 110021

  • 收稿日期:2007-09-24 修回日期:2008-01-29 出版日期:2008-05-10 发布日期:2008-05-10
  • 通讯作者: 苏来

Detection of DNA-protein crosslinks with modified comet assay

SU Lai, SONG Hong-Yu   

  1. National Research Center for New Drug Safety Evaluation, Shenyang 110021, China
  • Received:2007-09-24 Revised:2008-01-29 Online:2008-05-10 Published:2008-05-10
  • Contact: SU Lai

摘要:

将彗星实验进行改进以用于DNA-蛋白质交联作用的检测。利用甲醛对受试动物肝细胞的影响来判定此法是否适用于检测DNA-蛋白质交联。由于在实验中添加一定量的蛋白酶K, 可使单细胞在电泳时产生更大的迁移, 因此可以利用添加蛋白酶K前后的彗星尾距比来判断外来化合物对生物机体产生DNA损伤效应的时候是否有出现DNA-蛋白质交联作用。结果表明, 该方法快速、经济、灵敏度较高, 可以在单细胞水平对甲醛等强交联剂引发的不同组织的DNA-蛋白质交联效应进行检测, 希望该方法能成为指示DNA交联能力的有用工具。

关键词: 甲醛, 肝细胞, DNA断裂, DNA-蛋白质交联

Abstract:

Modifications of the comet assay were introduced to measure crosslinks and the effect of formaldehyde in liver cells of the tested animals was investigated by the new method to see whether the method is feasible. Since exposure of slides to proteinase K can increase DNA migration of the treated cells, the presence of DNA-protein crosslinks can be indicated by compare of the tail moment before and after the proteinase K added. The results showed that the modified protocol of the alkaline comet assay is a fast, inexpensive and sensitive tool for the detection of potent crosslinkers-induced DNA-protein crosslinks at single cell level. Due to its specific advantages, the modified comet assay seems to be a useful tool as a DNA crosslink potency indicator.