X-RAY-INDUCED DNA DOUBLE STRAND BREAK PRODUCTION AND REPAIR IN MAMMALIAN-CELLS AS MEASURED BY NEUTRAL FILTER ELUTION
X-RAY-INDUCED DNA DOUBLE STRAND BREAK PRODUCTION AND REPAIR IN MAMMALIAN-CELLS AS MEASURED BY NEUTRAL FILTER ELUTION
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DOI：
10.1093/nar/7.3.793
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发表时间：
1979-01-01
影响因子：
14.9
通讯作者：
KOHN, KW
中科院分区：
文献类型：
--
作者：
BRADLEY, MO;KOHN, KW
本文介绍了一种中性滤膜洗脱法检测X射线照射后小鼠L1210细胞DNA双链断裂的方法。该测定将检测由低至1000 rad的X射线诱导的双链断裂的数量。在中性条件下，通过过滤器的DNA洗脱速率随着X射线剂量的增加而增加。脱蛋白、pH和过滤器类型的某些条件显示出增加测定的灵敏度。过氧化氢和博莱霉素也诱导明显的DNA双链断裂，尽管双链与单链断裂的比例与X射线产生的比例不同。在中性条件下，通过在过滤器上裂解的DNA中引入HpA I限制性内切酶的双链切割导致快速洗脱，这意味着如果DNA中存在双链断裂，则该方法可以检测双链断裂。在氯化铯中洗脱的DNA带具有双链DNA标记。该证据表明，该检测试剂盒可检测DNA双链断裂。用5-10 krad X射线照射L1210细胞，可使大部分DNA双链断裂重新连接，半衰期约为40分钟。
This work presents a neutral filter elution method for detecting DNA double strand breaks in mouse L1210 cells after X-ray. The assay will detect the number of double strand breaks induced by as little as 1000 rad of X-ray. The rate of DNA elution through the filters under neutral conditions increases with X-ray dose. Certain conditions for deproteinization, pH, and filter type are shown to increase the assay's sensitivity. Hydrogen peroxide and Bleomycin also induce apparent DNA double strand breaks, although the ratios of double to single strand breaks vary from those produced by X-ray. The introduction of double strand cuts by HpA I restriction endonuclease in DNA lysed on filters results in a rapid rate of elution under neutral conditions, implying that the method can detect double strand breaks if they exist in the DNA. The eluted DNA bands with a double stranded DNA marker in cesium chloride. This evidence suggests that the assay detects DNA double strand breaks. L1210 cells are shown to rejoin most of the DNA double strand breaks induced by 5–10 krad of X-ray with a half-time of about 40 minutes.