Paul Moumita, Murray Vincent
School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, 2052, Australia.
Biomed Chromatogr. 2012 Mar;26(3):350-4. doi: 10.1002/bmc.1664. Epub 2011 Jun 16.
The determination of the sequence selectivity of DNA-damaging agents is very important in elucidating the mechanism of action of anti-tumour drugs. The development of automated capillary DNA sequencers with fluorescent labelling has enabled a more precise method for DNA sequence specificity analysis. In this work we utilized the ABI 3730 capillary sequencer with laser-induced fluorescence to examine the sequence selectivity of cisplatin with purified DNA sequences. The use of this automated machine enabled a higher degree of precision of both position and intensity of cisplatin-DNA adducts than previously possible with manual and automated slab gel procedures. A problem with artefact bands was overcome by ethanol precipitation. It was found that cisplatin strongly formed adducts with telomeric DNA sequences.
确定DNA损伤剂的序列选择性对于阐明抗肿瘤药物的作用机制非常重要。具有荧光标记的自动毛细管DNA测序仪的发展，使得DNA序列特异性分析有了更精确的方法。在这项工作中，我们利用带有激光诱导荧光的ABI 3730毛细管测序仪，来检测顺铂对纯化DNA序列的序列选择性。与之前的手动和自动平板凝胶程序相比，使用这种自动化仪器能使顺铂-DNA加合物的位置和强度具有更高的精度。通过乙醇沉淀克服了假带问题。研究发现，顺铂能与端粒DNA序列强烈形成加合物。
