^(32)P后标记新法研究DNA-苯醌加合物的生成  

BENZOQUINONE DNA ADDUCTS ANALYZED BY NUCLEASE P_1 MEDIATED ^(32)P-POSTLABELLING

作　　者：蒋湘宁[1] 刘淑芬[1] 徐晓白[1] 

机构地区：[1]中国科学院生态环境研究中心,北京100085

出　　处：《环境科学学报》1992年第4期407-414,共8页Acta Scientiae Circumstantiae

基　　金：国家自然科学基金资助项目

摘　　要：用以加P_1核酸酶提高灵敏度的^(32)P后标记新法研究了离体活细胞培养及试管反应形成的DNA-苯醌(BQ)加合物.4个被检加合物中有两个相应于苯醌与DNA上的胞嘧啶和鸟嘌呤作用形成的共价键加合物,其中较大的为首次发现的胞嘧啶-苯醌加合物.测得其相对标记率(RAL)试管条件为6.70×10^(-8)和8.93×10^(-9),细胞培养为8.23×10^(-9)和6.91×10^(-9),达到了在基因水平上研究DNA加合物的基本要求.With the super sensitivity, high reproducibility and accuracy, nuclease P1 mediated 32P-postlabelling version has been used successfully to analyze the DNA-BQ adducts formed from in vitro cultured cells, reaction of benzoquinone with calf thymus DNA and nucleoside monophposphates. It has been shown that the major one, which consists of more than 70% of the total radioactivities of DNA-BQ adducts detected, is from the deoxycytidine (dC) modified by benzoquinone while a minor one from deoxyguanosine (dG). The method is ca pable of detecting 1 adduct in 109-1010 DNA bases.

关 键 词：核酸酶 磷32 DNA-BQ 

分 类 号：Q3－3[生物学—遗传学]