改良碱性洗脱法可用于测定哺乳动物基因组DNA中完整的脱嘌呤位点。

Modified alkaline elution allows the measurement of intact apurinic sites in mammalian genomic DNA.

作者信息

Malvy C, Lefrançois M, Bertrand J R, Markovits J

机构信息

UMR 8532 CNRS, Physicochimie et Pharmacologie des Macromolécules Biologiques, Institut Gustave Roussy, rue Camille-Desmoulins, 94800, Villejuif, France.

出版信息

Biochimie. 2000 Aug;82(8):717-21. doi: 10.1016/s0300-9084(00)01152-4.

DOI:10.1016/s0300-9084(00)01152-4

PMID:11018287

Abstract

The presence of apurinic/apyrimidinic (AP) sites in cell genomes is known to be toxic and mutagenic. These lesions are therefore repaired in cells by efficient enzymatic systems. However, a report (Nakamura and Swenberg, Cancer Res. 59 (1999) 2522-2526) indicates an unexpected high rate of endogenous apurinic/apyrimidinic (AP) sites in genomic DNA in mammalian tissues. The technology used does not allow the authors to distinguish between intact AP sites and 3'cleaved AP sites. The corresponding values range between 2 and 4 sites per million of nucleotides in various human and rat tissues. Using a modified alkaline elution method we show here that the stationary level of intact AP sites is about 0.16 per million of nucleotides in leukemic mouse L1210 cells.

摘要

已知细胞基因组中脱嘌呤/脱嘧啶（AP）位点的存在具有毒性和致突变性。因此，这些损伤会在细胞中通过高效的酶系统进行修复。然而，一份报告（Nakamura和Swenberg，《癌症研究》59（1999）2522 - 2526）指出，哺乳动物组织基因组DNA中内源性脱嘌呤/脱嘧啶（AP）位点的发生率意外地高。作者所使用的技术无法区分完整的AP位点和3'切割的AP位点。在各种人类和大鼠组织中，相应的值在每百万个核苷酸中有2至4个位点。在此我们使用改良的碱性洗脱方法表明，白血病小鼠L1210细胞中完整AP位点的稳定水平约为每百万个核苷酸0.16个。