摘要
初步探索用聚合酶链反应(PCR)方法进行辐射诱导的线粒体DNA4977bp缺失分析的可行性。对正常人外周血进行2GyCoγ射线的照射,照射后提取包含线粒体DNA的全基因组DNA,用PCR方法进行线粒60体DNA4977bp缺失分析,同时扩增很少缺失的线粒体DNA片段(16S?ND1);进而比较照射前后线粒体DNA4977bp缺失水平。结果表明,用于分析线粒体DNA4977bp缺失的引物对在最佳试验条件下,可特异性扩增出所有2Gy照射诱导的线粒体DNA4977bp缺失;该扩增产物经纯化、测序和核苷酸同源性分析,证实线粒体缺失发生在线粒体DNA序列的8470?13446bp。用于扩增16S?ND1的引物对在很大的温度变动范围内,均能扩出目的DNA片段。所有外周血样品在照射前无线粒体DNA4977bp缺失,2Gy60Coγ射线照射后特异诱导出此缺失。说明用本研究中建立的PCR方法,可用来分析电离辐射诱导的外周血有核细胞线粒体DNA4977bp缺失水平。
This study is try to explore if the polymerase chain reaction (PCR) could be used to analyze the mitochondrial DNA 4977bp deletion (common deletion) induced by ionizing radiation. Peripheral blood samples were exposed to 2Gy Co γ-rays in vitro, and genomic DNA, including mitochondrial DNA, was extracted. A PCR 60 method was tried to analyze the common deletion, including seldom deleted 16S-ND1 gene. The common deletions before and after the 2Gy exposure were analyzed with the PCR method. The results showed that the common deletion PCR products were detected after exposure to 2Gy in each peripheral blood sample. The purified PCR products were sequenced and carried out BLAST on standard human mitochondrial genome sequence database. The result showed that the common deletion flanked between 8470bp and 13446bp. Before exposed to Co γ-rays, no common deletion 60 were not detected in the samples.These observations suggest that the PCR method, established in this study, could be used to detect the common deletion induced by ionizing radiation on peripheral lymphocytes samples.
出处
《辐射研究与辐射工艺学报》
CAS
CSCD
北大核心
2004年第5期311-314,共4页
Journal of Radiation Research and Radiation Processing
基金
国家人事部留学回国人员择优资助项目(2002年优秀类)
