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目的 :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 ,其产物经琼脂糖凝胶回收后与p GEM- 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 13 16 2 bp特大片段缺失突变。 结论 :此缺失是首次发现的 ,经测序证明为人线粒体 DNA最大片段的缺失。
Objective: To understand the mitochondrial DNA deletion mutation and its significance. Methods: Six kinds of methods were used to extract DNA from human peripheral blood. One method was to separate mitochondria and then extract mitochondrial DNA. The obtained human peripheral blood DNA as a template for PCR amplification, the product recovered by agarose gel p GEM-T vector connection, transformation, sequencing. Results: The results of electrophoresis showed that the PCR products of the six kinds of DNA extracted by PCR were the same, and the deletion of 13162 bp was found in mitochondrial DNA of human blood cells. CONCLUSION: This deletion was first discovered and was confirmed by sequencing as deletion of the largest segment of human mitochondrial DNA.