人外周血细胞线粒体DNA中存在特大片段缺失突变

了解线粒体ＤＮＡ大片段缺失突变及其意义。方法采用６种方法提取人外周血细胞ＤＮＡ,,其中１种方法先分离线粒体,再抽提线粒体ＤＮＡ。以得到的人外周血细胞ＤＮＡ为模板进行ＰＣＲ扩增,其产生经琼脂糖凝胶回收后与ｐＧＥＭ－Ｔ载体连接、转化、测序。结果６种方法抽提ＤＮＡ进行ＰＣＲ扩增后,产物电泳行为相同,发现人血细胞线粒体ＤＮＡ存在１３１６２ｂｐ特大片段缺失突变。结论此缺失首首发现的,经测序证明为人线粒体ＤＮ

A very large scale deletion mutation in human peripheral blood cells mitochondrial DNA

Objective:To understand large scale deletion mutation in mitochondrial DNA(mtDNA) and its significance. Methods: Six methods were used to extract human peripheral blood cells DNA of which one method was that mitochondria were isolated before mtDNA extraction. PCR was carried out using obtained human peripheral blood cells DNA as template. After extracted from agarose gels,PCR products was ligated with pGEM T vector, transformed and sequenced. Results: PCR products using template DNA extracted from above methods showed the same band in agarose electrophoresis, and a 13 162 bp mtDNA deletion mutation in human peripheral blood cells was found. Conclusion: This deletion is not reported previously and is the longest deletion in human mtDNA by sequencing.