| ^32P后标记法测DNA加合物 |
| |
| 引用本文： | 刘淑芬,蒋湘宁.^32P后标记法测DNA加合物[J].环境化学,1994,13(3):272-278. |
| |
| 作者姓名： | 刘淑芬 蒋湘宁 |
| |
| 作者单位： | 中国科学院生态环境研究中心!北京100085 |
| |
| 基金项目： | 国家自然科学基金资助项目 |
| |
| 摘 要： | ＾３２Ｐ后标记方法测ＤＮＡ－致癌物的加合物过程是用小球菌核酸酶和脾核酸酶将ＤＮＡ及加合物酶解成２′－脱氧核苷３′－单磷酸（３′－ｄＮｍＰ＋３′－ｄｘｍＰ）,以它为底物,用Ｔ４多核苷酸激酶催化,使γ＾３２ＰＡＴＰ的放射＾３２Ｐ转移反应到底物上,形成２′－脱氧核苷３′５′．ｄＮＤＰ＋３′５′＊ｄｘＤＰ）,用ＯＤＳ－ＴＬＣ和ＰＥＩ－ＴＬＣ结合将标记过的二磷酸分离分辨,用自显影制成加合物的指纹图,液闪计数
|
| 关 键 词： | 磷32后标记法 DNA加合物 测定 |
INVESTIGATION OF DNA-CARCINOGENS ADDUCTS BY ~(32)P-POSTLABELING |
| |
| Liu Shufen Jiang Xiangning.INVESTIGATION OF DNA-CARCINOGENS ADDUCTS BY ~(32)P-POSTLABELING[J].Environmental Chemistry,1994,13(3):272-278. |
| |
| Authors: | Liu Shufen Jiang Xiangning |
| |
| Abstract: | The 32P-postlabelimg procedure has been used to detect DNA-carcinogens adducts in vitro, in vivo, in human. Purified DNA was enzymatically digested to 2'-deoxyribonucleotide 3'-mono-phosphates (3'-dNmP, 3'-dxmP), Digested DNA (3'-dNmP) was converted into 32P-labeted 2'-deoxyribonucleatide-3'5'-bis-phosphate (3'5'"dNDP, 3'5'"dxDP) by incubation of 3'-dNmP with 32P ATP and T4 kinase. The adducts were separated by the chromatography (ODS-TLC and PEI-TLC) and were quantified by liquid scintillation counting. The sensitivity of 32P-postlabeling method was 1 adduct/1010 normal nucleotides. |
| |
| Keywords: | 32P-postlabeling procedure DNA adducts |
| 本文献已被 CNKI 维普 等数据库收录！ |
|
