摘要
^(32)P后标记方法测DNA-致癌物的加合物(in vitro,in vivo,in human样品)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T_4多核苷酸激酶催化,使γ^(32)P ATP的放射^(32)P转移反应到底物上,形成2’-脱氧核苷3’5’-二磷酸(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.液闪计数法计算出加合物浓度,灵敏度可达1加合物/10^(10)正常核酸.
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.
出处
《环境化学》
CAS
CSCD
北大核心
1994年第3期272-278,共7页
Environmental Chemistry
基金
国家自然科学基金资助项目
