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| 核酸序列水平定位DNA损伤的实验研究 | |
| 引用本文： | 高绪芳,饶朝龙,文卫华,衡正昌.核酸序列水平定位DNA损伤的实验研究[J].四川大学学报(医学版),2007,38(2):202-204,221. |
| 作者姓名： | 高绪芳 饶朝龙 文卫华 衡正昌 |
| 作者单位： | 1. 四川大学华西公共卫生学院,环境卫生学教研室,成都,610041;成都市疾病预防控制中心,卫生科 2. 成都中医药大学 3. 四川大学华西公共卫生学院,环境卫生学教研室,成都,610041 |
| 摘 要： | 目的 研究在核酸序列水平定位DNA损伤的方法.方法 培养TK6细胞,抽提基因组DNA,制备k-ras基因外显子2的单链探针.酶切基因组DNA构建损伤模型,应用依赖随机化末端连接物PCR(RDPCR)进行Southern blot,对产物进行测序.结果 酶切DNA的RDPCR产物在相应位置出现杂交条带,测序证实损伤位于HinfⅠ在k-ras外显子2的酶切位点.结论 将RDPCR和测序技术结合在一起,可在核酸水平上准确定位DNA损伤的碱基位置. |
| 关 键 词： | DNA损伤 定位检测 k-ras基因 依赖随机化末端连接物PCR 核酸序列 水平定位 损伤模型 实验 研究 Sequence Nucleic Acid DNA Lesion Location Research 碱基 准确定位 核酸水平 技术结合 酶切位点 基因外显子 杂交 位置 结果 测序 |
| 收稿时间： | 2006-05-22 |
| 修稿时间： | 2006-10-19 |
Experiment Research on the Location of DNA Lesion in the Nucleic Acid Sequence |
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| GAO Xu-fang,RAO Cao-long,WEN Wei-hua,HENG Zheng-cang.Experiment Research on the Location of DNA Lesion in the Nucleic Acid Sequence[J].Journal of West China University of Medical Sciences,2007,38(2):202-204,221. | |
| Authors: | GAO Xu-fang RAO Cao-long WEN Wei-hua HENG Zheng-cang |
| Institution: | Department of Environmental Health, West China School of Public Health, Sichuan University, Chengdu 610041, China. |
| Abstract: | OBJECTIVE: To research on establishing a method that can detect the DNA lesion at the level of nucleic acid. METHODS: The TK6 cell was cultured and treated, the genomic DNA was extracted, and the strand cleavage was induced by the endonuclease Hinf I. Then the genomic DNA was amplified by randomized terminal linker-dependent PCR (RDPCR), and hybridized by Southern hybridization with the single-stranded probes of the exon 2 of k-ras gene. The PCR products were sequenced. RESULTS: The clear hybridized band from the products of RDPCR was seen at the expectant position cut by Hinf I. The sequencing analysis showed that the position of DNA lesion linked by the linker was just the restriction site of Hinf I. CONCLUSION: It can be used to detect the DNA lesion at the level of nucleic acid sequence with the combination of sequence and RDPCR technologies. |
| Keywords: | DNA lesion Location detection k-ras gene RDPCR |
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