| 用^32P后标记方法测定典型有毒污染物的DNA加合物 |
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| 引用本文： | 刘淑芬,尹秀琴.用^32P后标记方法测定典型有毒污染物的DNA加合物[J].环境化学,1996,15(4):320-331. |
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| 作者姓名： | 刘淑芬 尹秀琴 |
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| 作者单位： | [1]中国科学院生态环境研究中心 [2]北京医科大学毒理系 |
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| 摘 要： | ＾３２Ｐ后标记方法测定生物样品里的有毒污染物－ＤＮＡ加合物的过程是用小球菌核酸酶和脾核酸酶将ＤＮＡ及加合物酶解成为２‘－脱氧核苷３’－单磷酸，以它为底物，在Ｔ４多核到激酶的催化下，与γ＾３２ＰＡＴＰ进行文笔磷的转移反应，形成带放射磷的２‘－脱氧核苷３’，５‘－二磷酸酯ＯＤＳ－ＴＬＣ吸附薄层板和ＰＥＩ－ＴＬＣ阴离子交换薄层板将标记过的带放射磷的＾＊３’，５‘－二磷酸酯分离分辨，然后用自显影技术制成加
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| 关 键 词： | 磷32后标记 DNA加合物 有毒污染物 测定 |
INVESTIGATION OF DNA-CARCINOGENS ADDUCT IN VITRO, IN VIVO, IN HUMAN BY 32P-POSTLABELING |
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| Liu Shufen Jiang Xiangning Xu Xiaobai.INVESTIGATION OF DNA-CARCINOGENS ADDUCT IN VITRO, IN VIVO, IN HUMAN BY 32P-POSTLABELING[J].Environmental Chemistry,1996,15(4):320-331. |
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| Authors: | Liu Shufen Jiang Xiangning Xu Xiaobai |
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| Abstract: | The 32P-postlabeling method used for detection DNA adduct was that DNA were enzymatically digested to deoxyribonucleoside 3 -monophosphates which were labeled to deoxyribonucleoside 3 5 -bisphosphates by T4 polynucleotide kinase catalyzed and r32P ATP. The labeled nucleotide were resolved on ODS-TLC and PEI-TLC, and detected by autoradiography. The sensitivity of method was 1 adduct per 1010 nucleotides. The 32P-postlabeling method was applied to analysis DNA adduct of styrene oxide, benzene, phenol, benzoquinone, 2-NF, DMA in vitro, in vivo, in human. A good correlation of DNA adduct and exposed concentration of chemical was found. |
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| Keywords: | DNA adduct 32P-postlabeIing |
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