cloned T₄ polynucleotide kinase

• 英文名称: cloned T₄ polynucleotide kinase

反应信息

• 作为试剂：
  描述：

  3-(2-Deoxy-β-D-erythro-pentofuranosyl)pyrimido[1,2-α]purin-10(3)-one-3'-monophosphate 在 sodium hydroxide 、 N,N-二羟乙基甘氨酸 、 disodium salt 、 cloned T₄ polynucleotide kinase 、 nuclease P₁ 、 sodium acetate 、 5’-三磷酸腺苷 、 zinc(II) chloride 作用下， 以 水 为溶剂， 反应 5.67h， 生成 3-(2-Deoxy-β-D-erythro-pentofuranosyl)pyrimido[1,2-α]purin-10(3)-one-5'-monophosphate
  参考文献：
  名称：

  An Improved ³²P-Postlabeling/High-Performance Liquid Chromatography Method for the Analysis of the Malondialdehye-Derived 1,N²-Propanodeoxyguanosine DNA Adduct in Animal and Human Tissues

  摘要：

  Malondialdehyde (MDA) is a major lipid peroxidation product that is mutagenic and tumorigenic. The MDA-modified DNA adduct, 3-(2-deoxy-beta-D-erythro-pentofuranosyl)pyr [1,2-alpha]purin-10(3H)-one (M(1)G), has been detected in human tissues and may be a marker of human cancer risk. In this paper, we describe an improved P-32-postlabeling/HPLC method for sensitive detection and quantitation of this MDA-modified 2'-deoxyribonucleotide adduct. Specific improvements include (i) unequivocal structural identification of the postlabeling products, both the 3',5'-bisphosphate of M(1)G (MDA-3',5'-dGDP) and the 5'-monophosphate of M1G (MDA-5'-dGMP); (ii) efficient separation of the P-32-postlabeling products by HPLC; and (iii) the incorporation of a synthetically prepared MDA-modified DNA (or the 3'-monophosphate of M(1)G) with a known modification level as an internal standard. This improved quantitative methodology provides high intra- and inter-assay reproducibility and has been applied to the analysis of this adduct in rodent and human samples.

  DOI：

  10.1021/tx9800497