使用自动测序仪确定DNA损伤的序列特异性。

Use of an automated sequencer to determine the sequence specificity of DNA damage.

作者信息

Hardman L C, Murray V

机构信息

School of Biochemistry and Molecular Genetics, University of New South Wales, Sydney, Australia.

出版信息

Biochem Mol Biol Int. 1997 Jun;42(2):349-59. doi: 10.1080/15216549700202751.

DOI:10.1080/15216549700202751

PMID:9238534

Abstract

An automated sequencer was used to determine the sequence specificity of DNA damage caused by hedamycin in the plasmid pUC19 using a linear amplification/Taq DNA polymerase method. Previously, manual DNA sequencers have been in widespread use to investigate the sequence specificity of a DNA damaging agent. Manual DNA sequencers are restricted in the length of DNA sequence that can be read at base pair resolution for densitometry. An automated sequencer can greatly expand on the length of analysable DNA sequence. An additional important capability of the automated sequencer, is the ability to quantitate the intensity of damage at each base pair site. Thus we have used the automated sequencer to elucidate the sequence specificity of DNA damage for 300 bp. We have carried out an extended analysis of the sequence specificity of hedamycin DNA damage and found that the sequence 5'-cGt-3', tGt and cGg are preferentially damaged. The sequence specificity of cisplatin was also investigated.

摘要

使用线性扩增/Taq DNA聚合酶方法，通过自动测序仪来确定柔红霉素在质粒pUC19中引起的DNA损伤的序列特异性。以前，手动DNA测序仪已被广泛用于研究DNA损伤剂的序列特异性。手动DNA测序仪在可通过光密度法以碱基对分辨率读取的DNA序列长度方面受到限制。自动测序仪可以大大扩展可分析DNA序列的长度。自动测序仪的另一个重要功能是能够定量每个碱基对位点的损伤强度。因此，我们使用自动测序仪阐明了300 bp的DNA损伤的序列特异性。我们对柔红霉素DNA损伤的序列特异性进行了扩展分析，发现序列5'-cGt-3'、tGt和cGg优先受损。还研究了顺铂的序列特异性。