SNP分型结合甲基化亲源相关性及与暴力犯罪行为的相关性研究

发布时间：2018-04-28 17:43

本文选题：法医物证学 + 单核苷酸多态性　；参考：《复旦大学》2013年硕士论文

【摘要】：目的建立经典变性梯度凝胶电泳(denaturing gradient gel electrophoresis, DGGE)技术与新发展的焦磷酸测序技术(pyrosequencing)对小核核糖核蛋白多肽N基因(small nuclear ribonucleoprotein polypeptide N,SNRPN) SNP位点rs220030位点的分型方法,比较焦磷酸测序技术在SNP分型上相对DGGE电泳技术的优势；建立应用焦磷酸测序技术分析CpG甲基化状态的方法,探讨SNP位点rs220030用于亲源等位基因判定的可行性。方法应用DGGE技术对97例上海地区汉族家系样本rs220030位点进行分型,同时应用焦磷酸测序技术对其中25例家系样本的rs220030位点再次分型,并对两种方法在SNP分型上进行比较；通过重亚硫酸盐修饰联合焦磷酸测序技术分析随机的2组家系rs220030位点上游CpG甲基化状态,判断甲基化是否有亲源相关性。结果经DGGE电泳检测97例家系样本SNP位点rs220030分型结果为C纯合子20例,T纯合子29例,C/T杂合子48例；经焦磷酸测序检测25例样本结果与DGGE检测结果一致；经焦磷酸测序技术分析,2组家系子代的rs220030位点上游CpG甲基化状态均与母亲较相似。结论相比经典DGGE技术,焦磷酸测序技术更精确、更方便,适合大样本、高通量SNP分型；重亚硫酸盐修饰联合焦磷酸测序技术可以精确分析CpG甲基化状；rs220030有用于亲源等位基因判定的可行性。目的探讨GABRB3基因多态性与暴力犯罪行为之间是否存在相关性,筛选出GABRB3基因启动子区可能与暴力犯罪行为存在相关性的SNP位点；并根据有暴力犯罪前科人群中所检测出的GABRB3基因启动子区SNP位点多态性情况,探讨GABRB3基因启动子区多态性与表达之间的关系。方法采用PCR-直接测序的方法对150例有暴力犯罪行为前科的中国汉族男性的DNA样本进行GABRB3基因待测SNP位点检测,以生物信息数据库中提供的中国正常健康汉族男性人群的群体遗传学信息(包括中国北京汉族人群、中国南方汉族人群以及两者合计)作为正常对照组,运用卡方检验(P为0.05)进行三次交叉对比,分析暴力犯罪组与正常人群相比在GABRB3基因多态性频率分布是否有统计学差异。结果待测SNP点选择为rs20317、rs8179184、rs4906902;其中rs20317在三次交叉对比的结果P均0.05；而rs8179184、rs4906902在三次交叉对比结果P均0.05；同一个样本rs4906902与rs8179184两者基因型相关联；结论rs20317中C等位基因可能与暴力犯罪行为有相关性,可能能作为暴力犯罪风险预防基因；GABRB3启动子区SNP位点多态性对基因转录表达造成影响,进而影响其功能,可能引起行为学变化；GABRB3基因启动子区内SNP位点rs4906902与rs8179184存在紧密连锁现象。
[Abstract]:Objective to establish a method for typing the rs220030 site of small ribonucleoprotein polypeptide N gene, small nuclear ribonucleoprotein polypeptide N, SNRPN, by denaturing gradient gel electrophoresis, DGGE) and pyrosequencing. To compare the advantages of pyrosequencing over DGGE electrophoresis in SNP typing, to establish a method to analyze the methylation status of CpG by pyrosequencing, and to explore the feasibility of using SNP locus rs220030 in the determination of parent alleles. Methods the rs220030 loci of 97 families of Han nationality in Shanghai were classified by DGGE and 25 of them were classified by pyrosequencing. The SNP genotyping was compared between the two methods. The methylation status of CpG in the upstream of rs220030 locus in two families was analyzed by double sulfite modification combined with pyrosequencing to determine whether the methylation had a positive correlation. Results the results of rs220030 typing of SNP loci in 97 pedigree samples by DGGE electrophoresis were as follows: C homozygote 20 cases, T homozygote 29 cases, C / T heterozygote 48 cases, pyrosequencing was consistent with DGGE test in 25 cases, C homozygote was detected in 20 cases and T homozygote in 29 cases. The CpG methylation status in the upstream of rs220030 locus was analyzed by pyrosequencing technique in the two families, which was similar to that of the mother. Conclusion compared with classical DGGE, pyrosequencing is more accurate, convenient and suitable for large sample and high throughput SNP typing. Double sulfite modification combined with pyrosequencing can accurately analyze the feasibility of CpG methylation rs220030 for the determination of parent alleles. Objective to investigate the correlation between GABRB3 gene polymorphism and violent criminal behavior, and to screen out the SNP locus in the promoter region of GABRB3 gene that may be associated with violent criminal behavior. The relationship between the polymorphism and expression of GABRB3 gene promoter region was discussed according to the polymorphism of SNP locus in the promoter region of GABRB3 gene in the population with criminal record of violent crime. Methods the SNP loci of GABRB3 gene were detected in 150 DNA samples of Chinese Han nationality men with criminal record of violence by PCR- direct sequencing. The population genetic information (including the Han population in Beijing, the Han population in the south of China and the two groups) provided in the bioinformatics database was used as the normal control group. Chi-square test (P = 0.05) was used to analyze whether the frequency distribution of GABRB3 gene polymorphism in the violent crime group was statistically different from that in the normal population. Results the selection of SNP sites to be tested was rs203179184 / rs4906902, in which the results of rs20317 were all 0.05 in three cross contrasts, while the results of rs8179184rs4906902 were all 0.05 in three cross contrasts. The genotype of rs4906902 and rs8179184 in the same sample was correlated with that of rs8179184. Conclusion the C allele in rs20317 may be associated with violent criminal behavior, and it may be used as the SNP locus of GABRB3 promoter to influence the transcription and expression of GABRB3 gene, and then affect the function of GABRB3 gene. It may cause behavioral changes. There is a close linkage between rs4906902 and rs8179184 in the promoter region of GABRB3 gene.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：D919

【参考文献】