RAPID ISOLATION, HYDROLYSIS AND CHROMATOGRAPHY OF FORMALDEHYDE-MODIFIED DNA
RAPID ISOLATION, HYDROLYSIS AND CHROMATOGRAPHY OF FORMALDEHYDE-MODIFIED DNA
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DOI：
10.1016/0378-4347(84)80202-9
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发表时间：
1984-01-01
期刊：
影响因子：
--
通讯作者：
HEFLICH, RH
中科院分区：
文献类型：
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作者：
BELAND, FA;FULLERTON, NF;HEFLICH, RH
脱氧腺苷、脱氧鸟苷、脱氧胞苷和胸苷与甲醛反应。高效液相色谱（HPLC）分析表明，每种脱氧核苷形成1种主要产物。除胸苷产物外，这些加合物通过NMR光谱分析并鉴定为环外胺的羟甲基衍生物。小牛胸腺DNA与[3 H]甲醛孵育，纯化后，酶水解为核苷。HPLC分析表明存在相当大比例的非共价结合的甲醛和以下羟甲基加合物，按浓度递减的顺序列出：N6-羟甲基脱氧腺苷. mchgt。N4-羟甲基脱氧胞苷> N2-羟甲基脱氧鸟苷。用[3 H]甲醛孵育中国仓鼠卵巢（CHO）细胞，导致甲醛代谢掺入嘌呤和嘧啶中，以及与DNA非共价结合的可观浓度的甲醛。HPLC分析清楚地表明CHO细胞基因组中存在N6-羟甲基脱氧腺苷。
Deoxyadenosine, deoxyguanosine, deoxycytidine and thymidine were reacted with formaldehyde. High-performance liquid chromatographic (HPLC) analysis indicated that each deoxynucleoside had formed 1 major product. With the exception of the thymidine product, these adducts were analyzed by NMR spectroscopy and identified as hydroxymethyl derivatives at the exocyclic amines. Calf thymus DNA was incubated with [3H]formaldehyde and, after purification, enzymatically hydrolyzed to nucleosides. HPLC analysis indicated the presence of a substantial proportion of noncovalently bound formaldehyde and the following hydroxymethyl adducts, listed in order of decreasing concentration: N6-hydroxymethyldeoxyadenosine .mchgt. N4-hydroxymethyldeoxycytidine > N2-hydroxymethyldeoxyguanosine. Incubation of Chinese hamster ovary (CHO) cells with [3H]formaldehyde resulted in metabolic incorporation of the formaldehyde into purines and pyrimidines plus an appreciable concentration of formaldehyde noncovalently associated with the DNA. HPLC analysis clearly indicated the presence of N6-hydroxymethyldeoxyadenosine in the CHO cell genome.