本文選題：法醫(yī)物證學 + 單核苷酸多態(tài)性��； 參考：《復旦大學》2013年碩士論文

【摘要】：目的建立經(jīng)典變性梯度凝膠電泳(denaturing gradient gel electrophoresis, DGGE)技術與新發(fā)展的焦磷酸測序技術(pyrosequencing)對小核核糖核蛋白多肽N基因(small nuclear ribonucleoprotein polypeptide N,SNRPN) SNP位點rs220030位點的分型方法,比較焦磷酸測序技術在SNP分型上相對DGGE電泳技術的優(yōu)勢；建立應用焦磷酸測序技術分析CpG甲基化狀態(tài)的方法,探討SNP位點rs220030用于親源等位基因判定的可行性。方法應用DGGE技術對97例上海地區(qū)漢族家系樣本rs220030位點進行分型,同時應用焦磷酸測序技術對其中25例家系樣本的rs220030位點再次分型,并對兩種方法在SNP分型上進行比較；通過重亞硫酸鹽修飾聯(lián)合焦磷酸測序技術分析隨機的2組家系rs220030位點上游CpG甲基化狀態(tài),判斷甲基化是否有親源相關性。結果經(jīng)DGGE電泳檢測97例家系樣本SNP位點rs220030分型結果為C純合子20例,T純合子29例,C/T雜合子48例；經(jīng)焦磷酸測序檢測25例樣本結果與DGGE檢測結果一致；經(jīng)焦磷酸測序技術分析,2組家系子代的rs220030位點上游CpG甲基化狀態(tài)均與母親較相似。結論相比經(jīng)典DGGE技術,焦磷酸測序技術更精確、更方便,適合大樣本、高通量SNP分型；重亞硫酸鹽修飾聯(lián)合焦磷酸測序技術可以精確分析CpG甲基化狀；rs220030有用于親源等位基因判定的可行性。 目的探討GABRB3基因多態(tài)性與暴力犯罪行為之間是否存在相關性,篩選出GABRB3基因啟動子區(qū)可能與暴力犯罪行為存在相關性的SNP位點；并根據(jù)有暴力犯罪前科人群中所檢測出的GABRB3基因啟動子區(qū)SNP位點多態(tài)性情況,探討GABRB3基因啟動子區(qū)多態(tài)性與表達之間的關系。方法采用PCR-直接測序的方法對150例有暴力犯罪行為前科的中國漢族男性的DNA樣本進行GABRB3基因待測SNP位點檢測,以生物信息數(shù)據(jù)庫中提供的中國正常健康漢族男性人群的群體遺傳學信息(包括中國北京漢族人群、中國南方漢族人群以及兩者合計)作為正常對照組,運用卡方檢驗(P為0.05)進行三次交叉對比,分析暴力犯罪組與正常人群相比在GABRB3基因多態(tài)性頻率分布是否有統(tǒng)計學差異。結果待測SNP點選擇為rs20317、rs8179184、rs4906902;其中rs20317在三次交叉對比的結果P均0.05；而rs8179184、rs4906902在三次交叉對比結果P均0.05；同一個樣本rs4906902與rs8179184兩者基因型相關聯(lián)；結論rs20317中C等位基因可能與暴力犯罪行為有相關性,可能能作為暴力犯罪風險預防基因；GABRB3啟動子區(qū)SNP位點多態(tài)性對基因轉錄表達造成影響,進而影響其功能,可能引起行為學變化；GABRB3基因啟動子區(qū)內(nèi)SNP位點rs4906902與rs8179184存在緊密連鎖現(xiàn)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.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：D919

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相關期刊論文 前1條

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本文編號：1816251