A study of the pigment which inhibits enzymatic reaction

抑制酶反应色素的研究

• 批准号: 14570397
• 负责人: KUROSAKI Kunihiko
• 金额: $ 1.54万
• 依托单位: Tokyo Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570397/
• 关键词: DNA typing; pigment; hair dye; 染毛剤; 染色剤; DNA多型; PCR

When some pigment infiltrate into a forensic or an anthropological sample such as a hair and a human bone, it is known that DNA typing is impossible in many cases because PCR reaction does not advance. I carried out this study for the purpose of establishing methods for DNA typing from the sample in which the pigment was included.At first, I tried to remove the pigment of hair dye which inhibits PCR reaction, but could not unfortunately find effective methods to do it. However, even in the case of a hair treated with perpetual hair dye, it became clear that PCR is successful by using template DNAs extracted by the method without the dissolution of the pilar cortex. On the other hand, I paid my attention to natural pigment, which inhibited PCR, in the burial soil. Then, I tried to extract DNA from ancient bone without enzyme inhibition by these pigment ingredients. As a result, even in DNA extracted from the dyed bones, a success rate of DNA typing could be raised by my original method of DNA extraction using silica powder.In this study, I succeeded in overcoming most, of the enzyme inhibition with natural and synthetic pigments.

当某些色素渗入法医学或人类学样品如头发和人骨中时，已知在许多情况下DNA分型是不可能的，因为PCR反应不能进行。本研究的目的是从含有色素的样品中建立DNA分型的方法。起初，我试图去除抑制PCR反应的染发剂色素，但不幸的是，我无法找到有效的方法。然而，即使在用永久染发剂处理的头发的情况下，很明显，通过使用由该方法提取的模板DNA而不溶解皮拉尔皮质，PCR是成功的。另一方面，我注意到了埋藏土壤中抑制PCR的天然色素。然后，我尝试从古骨骼中提取DNA，而不受这些色素成分的酶抑制。结果，即使是从染色的骨头中提取的DNA，也可以通过使用二氧化硅粉末的原始DNA提取方法提高DNA分型的成功率。在本研究中，我成功地克服了天然和合成色素对酶的大部分抑制。