本文選題：線粒體 + DNA(mtDNA)　； 參考：《山西醫(yī)科大學(xué)》2002年碩士論文

【摘要】： 目的 獲得線粒體DNA HVI在太原地區(qū)漢族群體中分布的序列多態(tài)性數(shù)據(jù)，以探討線粒體DNA序列多態(tài)性在法醫(yī)學(xué)個(gè)體識別中應(yīng)用的理論基礎(chǔ)。并探索采用PCR-SSCP技術(shù)作為一種檢測線粒體DNA序列多態(tài)性的篩選方法。 方法 隨機(jī)抽取58名太原地區(qū)漢族群體無血緣關(guān)系個(gè)體的靜脈血，EDTA抗凝，改良的TKM法提取mtDNA，PCR擴(kuò)增、純化后，應(yīng)用377序列分析儀進(jìn)行直接測序分析；對毛發(fā)、指甲、骨骼等微量或嚴(yán)重降解檢材進(jìn)行mtDNA序列分析；采集同一尸體血液、肌肉、肝、腎、心肌、不同部位的毛干等組織進(jìn)行同一性檢測；對20個(gè)兩代家系進(jìn)行突變觀察。將上述58名無關(guān)個(gè)體的mtDNA擴(kuò)增后，，10％非變性聚丙烯酰胺凝膠(含10％甘油)電泳，銀染，進(jìn)行SSCP分析。 結(jié)果 在線粒體DNAnt16081-16546區(qū)間發(fā)現(xiàn)有55種單倍型，其中53種是唯一的，另2種分別為2個(gè)和3個(gè)個(gè)體所共有。該區(qū)間共有74處堿基變異，其可變堿基的變異形式主要為堿基替換(轉(zhuǎn)換和顛換)、插入和缺失；堿基轉(zhuǎn)換(77.0％)明顯高于顛換(9.46％)、插入(12.2％)、缺失(1.35％)。該區(qū)段基因變異度為99.8％，偶合概率為1.96％。微量血痕、毛發(fā)、指甲、骨骼進(jìn)行 山百醫(yī)科大學(xué) 2002月卜祠吐士研究生畢習(xí)匕自侖文 mtDNA序列分析均能準(zhǔn)確測定堿基序列。人體不同組織序列一致 性檢測中觀察到頭前部的一根毛發(fā)mtDNA序列16249處堿基N 為G/A，而其余組織此處堿基為A。家系突變觀察結(jié)果顯示一對 母親一孩子序列不一致，母親為16193.ZC，孩子為16193.C。SSCP 分析結(jié)果在同一張凝膠上序列不同的8個(gè)樣品，可平均檢出5 種帶型，檢出率可達(dá)62.5%。 結(jié)論所檢測區(qū)間ntl6os一16546位于線粒體DNAHVI，從 基因變異度和偶合概率兩項(xiàng)指標(biāo)，及mtONA可對那些ONA嚴(yán)重 降解或不能提供足夠核ONA的檢材進(jìn)行分析，顯示mtDNA可作 為良好的遺傳標(biāo)記進(jìn)行個(gè)體識別。與不同人群比較結(jié)果中發(fā)現(xiàn)四 個(gè)漢族群體間差異無顯著性，而與瑞士高加索人群都存在顯著性 差異。人體不同組織一致性檢驗(yàn)結(jié)果和家系突變中觀察到異質(zhì) 型，異質(zhì)型的出現(xiàn)并不會(huì)使mtDNA的法醫(yī)學(xué)分析失效，但應(yīng)考慮 它對判定結(jié)果的影響。本文應(yīng)用的熒光染料標(biāo)記全自動(dòng)測序技術(shù) 具有準(zhǔn)確性高，序列結(jié)果穩(wěn)定;靈敏度高;電泳分離自動(dòng)化等特 點(diǎn)。但同時(shí)也存在一些不足，如測序需多次擴(kuò)增，容易污染;該 方法費(fèi)用昂貴，需要特殊的儀器等。SSCP分析技術(shù)實(shí)驗(yàn)方法簡 單、易操作;實(shí)驗(yàn)成本低，可作為一種篩選大量嫌疑檢材的方法。
[Abstract]:Objective to obtain the sequence polymorphism data of mitochondrial DNA HVI distribution in Taiyuan Han population and to explore the theoretical basis for the application of mitochondrial DNA polymorphism in forensic individual identification. PCR-SSCP was used as a screening method to detect mitochondrial DNA polymorphism. Methods in 58 unrelated individuals of Han nationality in Taiyuan area, the anticoagulant mtDNA-EDTA was extracted by modified TKM method. After purification, direct sequencing was performed by 377 sequence analyzers, and hair and fingernails were analyzed. Trace or severe degradation of bone samples were analyzed by mtDNA sequence, blood, muscle, liver, kidney, myocardium, hair stem of different parts of the same cadaver were collected for identity detection, and 20 generations of families were observed for mutation. After mtDNA amplification, 10% non-denatured polyacrylamide gel (including 10% glycerol) was detected by silver staining and SSCP analysis. Results there were 55 haplotypes in the mitochondrial DNAnt16081-16546 region, of which 53 were unique and the other two were shared by 2 and 3 individuals, respectively. There are 74 base variations in this region, and the variable bases are mainly in the form of base substitution (transformation and transversion, insertion and deletion; base conversion 77.0), which is obviously higher than that of 9.46%, with the insertion of 12. 2% and the absence of 1.35%. The genetic variation of this region is 99.8 and the probability of coupling is 1.96. Traces of blood, hair, nails, bones Shanbai Medical University In February 2002, the graduate student of Bushi Tutu graduated from Zilun. MtDNA sequence analysis can accurately determine the base sequence. The sequence of different tissues in human body is consistent. A mtDNA sequence of 16249 base N was observed in a hair in the anterior part of the head. G / A, and the rest of the tissues have a base A. Pedigree mutation observation showed a pair of Mother one child sequence is inconsistent, mother is 16193. ZC, child is 16193.C.SSCP Eight samples with different sequences on the same gel can be detected with an average of 5. 5% The detection rate was 62.5%. Conclusion the detected interval ntl6os-16546 is located in the mitochondrial DNA HVI. Genetic variability and coupling probability, and mtONA can be used to treat those ONA seriously. Degradation or failure to provide sufficient nuclear ONA samples for analysis, indicating that mtDNA can be used To identify individuals for good genetic markers. In comparison with different population groups, we find that There was no significant difference between Han population and Swiss Caucasian population. Difference. Heterogeneity was observed in the results of consistency test of different human tissues and family mutation. The appearance of heterogeneity does not invalidate the forensic analysis of mtDNA, but it should be considered Its effect on the result. Automatic sequencing technique for fluorescent dye labeling in this paper High accuracy, stable sequence results, high sensitivity; Electrophoretic separation automation, etc. Point. But at the same time there are some shortcomings, such as sequencing needs multiple amplification, easy to pollute; The method is expensive and requires special instruments and so on. The method of SSCP analysis is simple. Simple, easy to operate, low cost, can be used as a method to screen a large number of suspected materials.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2002
【分類號】：D919

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 趙永斌;于長春;周慧;;漢族起源與發(fā)展的遺傳學(xué)探索[J];吉林師范大學(xué)學(xué)報(bào)(自然科學(xué)版);2012年04期

相關(guān)碩士學(xué)位論文 前1條

1 鄧晨;基于磁性微粒純化法醫(yī)樣本DNA方法的建立[D];西北大學(xué);2012年

本文編號：1873800