[曲拉通X-100处理对大鼠肝细胞核DNA完整性及DNA:RNA:蛋白质比率的影响]

[Effect of treatment with Triton X-100 on DNA integrity and DNA:RNA: protein ratio of rat liver nuclei].

作者信息

Carlo P, Martelli A, Bignone F A

出版信息

Boll Soc Ital Biol Sper. 1981 Nov 30;57(22):2203-8.

Abstract

A procedure is described that gives clean nuclei with intact DNA from a rat liver cell suspension. Cytoplasmic contamination is removed by successive treatments with a non ionic detergent, Triton X-100 (0.75%, v/v). We found that with ratios DNA:RNA: protein of 1:0.09:3.29 (2 Triton X-100 steps) the integrity of DNA is preserved: further decrease in RNA and protein content (3 Triton X-100 steps) causes DNA breakage, probably because of extraction of nuclear proteins. In order to estimate DNA integrity, its viscosity was determined by the use of a new oscillating crucible viscometer; this method makes possible the evaluation of extremely small levels of DNA damage as that induced by 0.067 mg/kg of DMNA injected i.p.

摘要

本文描述了一种从大鼠肝细胞悬液中获取具有完整DNA的纯净细胞核的方法。通过用非离子去污剂Triton X-100（0.75%，v/v）连续处理来去除细胞质污染。我们发现，当DNA:RNA:蛋白质的比例为1:0.09:3.29（经过两步Triton X-100处理）时，DNA的完整性得以保留：RNA和蛋白质含量进一步降低（经过三步Triton X-100处理）会导致DNA断裂，这可能是由于核蛋白的提取所致。为了评估DNA的完整性，使用一种新型振荡坩埚粘度计测定其粘度；这种方法能够评估极低水平的DNA损伤，如腹腔注射0.067 mg/kg的二甲基亚硝胺（DMNA）所诱导的损伤。