头发mtDNA4977bp缺失与年龄的关系研究

发布时间：2018-06-08 06:56

本文选题：线粒体 + 头发　；参考：《重庆医科大学》2006年硕士论文

【摘要】： 目的:研究头发mtDNA4977bp片段缺失与人群年龄的关系,建立相关方程,以期在微量物证中寻找更多的信息,为分子水平构建犯罪分子特征提供更多的可能。方法:按中国人口自然分布规律,随机剪取54名汉族健康志愿者的头发若干,各取4根用无水乙醇清洗、蒸馏水漂洗后,自然风干,从近毛囊部位截取1.5cm,剪成0.3cm左右的片段,放入0.5ml EP管中,加chelex-100和蛋白酶K消化抽提mtDNA,用所设计的引物分别对mtDNA保守区和4977bp缺失突变区进行聚合酶链反应(PCR),若4977bp处不发生缺失,常规PCR不能扩增正常约5000bp的碱基,所以没有产物,反之,可以扩增出线粒体片段缺失后相应遗留序列的PCR产物。将扩增产物经琼脂糖凝胶电泳分离,EB染色后,在Bio-Rad紫外凝胶成像仪扫描成像,确定其激光密度。以保守内参照区扩增产物激光密度作为总量,缺失区扩增产物激光密度值作为缺失量,用缺失量/总量计算缺失率。结果:25岁以下几乎无缺失,缺失区很少扩增出产物,大于25岁者缺失区逐渐可扩增出产物,且随年龄增长,相同条件下扩增产物增多,4977bp缺失率增大,各年龄段组间比较均P0.05。根据统计分析,缺失率与年龄有直线相关趋势,相关系数r=0.98,可以建立相关回归方程Age=0.74(±0.02)×dmtDNA+21.03(±0.80)(dmtDNA为缺失
[Abstract]:Objective: To study the relationship between the loss of mtDNA4977bp fragments of hair and the age of the population and establish the correlation equation, in order to find more information in the trace evidence, and to provide more possibilities for the construction of criminal characteristics at the molecular level.
Methods: according to the natural distribution law of Chinese population, the hair of 54 Han healthy volunteers was cut randomly, each 4 was washed with anhydrous ethanol. After the distilled water was rinsed, the natural wind was dried, the 1.5cm was cut from the near hair follicle, and the fragments were cut into about 0.3cm. In the 0.5ml EP tube, the mtDNA was digested with Chelex-100 and protease K, and the designed citation was used. Polymerase chain reaction (PCR) was performed on the mtDNA conservative region and the 4977bp deletion mutation region respectively. If no deletion occurred at 4977bp, the conventional PCR could not amplify the normal base of approximately 5000bp, so there was no product. On the contrary, the PCR products of the corresponding remnants of the mitochondrial fragment could be amplified. The amplified products were separated by agarose gel electrophoresis and EB. After staining, the laser density was determined by scanning the Bio-Rad ultraviolet gel imaging instrument. The laser density of the amplified product in the internal reference area was taken as the total amount, the laser density value of the amplified product in the missing region was used as the missing amount, and the loss rate was calculated by the amount of loss / total.
Results: there was almost no deletion under 25 years of age, and there was little amplification of the product in the missing area, and the product was gradually amplified in the missing area of the people over 25 years old, and with the increase of age, the amplification products increased and the loss rate of 4977bp increased. The correlation coefficient of the loss rate was linearly correlated with age, and the correlation coefficient was r=0.98, as compared with the statistical analysis of P0.05.. Correlation regression equation Age=0.74 (+ 0.02) x dmtDNA+21.03 (+ 0.80) can be established (dmtDNA is missing).
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：D919

【参考文献】