Complete mitochondrial genome sequences for 265 African American and U.S. “Hispanic” individuals

Entire mitochondrial genome (mtGenome) sequences of 265 unrelated African American and U.S. “Hispanic” individuals were generated.     

mtDNA测序结果与Anderson标准序列的比对－ClustalX等共享软件在法医物证学上的应用

目的 对多个样本的线粒体DNA(mtDNA)高变区测序结果与Anderson标准序列进行比对分析。方法 利用ABI测序仪测定生物学样本的mtDNA高变区序列,得到测序结果文件,通过Chromas、 SeqVerter软件将之转换为aln文件,用ClustalX软件与Anderson标准序列(txt文件)进行比对,确定突变点的碱基排列次序和位置。结果Chromas、SeqVerter和ClustalX软件界面友好,操作简便,可以方便地用于多个样本DNA序列的比较,结果直观,易于判读。结论　运用Chromas、SeqVerter和ClustalX等共享软件,可成功地对多个样本的mtDNA高变区序列进行比对分析。     

中国新疆喀什维吾尔族群体mtDNA D环区多态性

<正> 为更好地应用mtDNA序列多态性进行个体识别和亲缘关系鉴定,本文应用PCR产物直接测序法研究99名新疆喀什维族无关个体mtDNA D环区全序列特征,并对其进行统计分析,意在建立mtDNA多态性应用的理论根据。     

中国藏族群体mtDNA控制区多态性

<正> 对不同民族人群进行mtDNA序列分析,可以揭示不同群体mtDNA多态性特点,有助于mtDNA分析技术的法医学应用。本文作者用PCR产物直接测序法,对中国西藏那曲地区藏族42名无关个体的mtDNA控制区进行全序列测定,并对其进行统计学分析。     

Mitochondrial DNA control region population data from Macedonia

Mitochondrial DNA sequences of the entire control region were analyzed in 200 unrelated individuals from Macedonia. A total of 163 different haplotypes were found as determined by 177 polymorphic sites. The probability of a random match was calculated as 1:121 (0.83%). The basic phylogenetic structure of the Macedonian population as derived from its haplogroup distribution is in agreement with other West-Eurasian populations. Upon publication, the population data are going to be available in the EMPOP database (www.empop.org) [W. Parson, A. Dür, EMPOP—a forensic mtDNA database, FSI:Genetics 1 (2) (2007) 88–92; W. Parson, A. Brandstätter, A. Alonso, N. Brandt, B. Brinkmann, A. Carracedo, et al., The EDNAP mitochondrial DNA population database (EMPOP) collaborative exercises: organisation, results and perspectives, Forensic Sci. Int. 139 (2–3) (2004) 215–226.].     

mtDNA—HVⅠ和细胞色素b片段的复合扩增及其法医学应用

目的探讨复合扩增mtDNA D环HV I和细胞色素b片段进行种属鉴定和个体识别的方法及mtDNA-HV I多态性。方法用两对引物同步扩增HV I片段与细胞色素b片段,银染显带检测扩增产物,ABI377测序仪及荧光测序技术分析扩增产物序列多态性。结果人类有279bp,358bp两条带,动物只有358bp一条带。通过对131例随机广东汉族人群个体进行mtDNA控制区(15997～16236))序列测定统计,得出此区域的序列多态性。共发现69个位点变异,平均每个个体存在2.679个碱基突变,检出67个单倍型,基因多样性为97.92％。结论mtDNA控制区(15997—16236)具有较高的序列多态性。为良好的个体识别标记。复合扩增mtDNA D环HV I与细胞色素b片段进行测序分析可以同步进行种属鉴定和个体识别。     

Typing of mitochondrial DNA coding region SNPs of forensic and anthropological interest using SNaPshot minisequencing

The development of new methodologies for high-throughput SNP analysis is one of the most stimulating areas in genetic research. Here, we describe a rapid and robust assay to simultaneously genotype 17 mitochondrial DNA (mtDNA) coding region SNPs by minisequencing using SNaPshot. SNaPshot is a methodology based on a single base extension of an unlabeled oligonucleotide with labeled dideoxy terminators. The set of SNPs implemented in this multiplexed SNaPshot reaction allow us to allocate common mitochondrial West Eurasian haplotypes into their corresponding branch in the mtDNA skeleton, with special focus on those haplogroups lacking unambiguous diagnostic positions in the first and second hypervariable regions (HVS-I/II; by far, the most common segments analyzed by sequencing). Particularly interesting is the set of SNPs that subdivide haplogroup H; the most frequent haplogroup in Europe (40–50%) and one of the most poorly characterized phylogenetically in the HVS-I/II region. In addition, the polymorphic positions selected for this multiplex reaction increase considerably the discrimination power of current mitochondrial analysis in the forensic field and can also be used as a rapid screening tool prior to full sequencing analysis. The method has been validated in a sample of 266 individuals and shows high accuracy and robustness avoiding both the use of alternative time-consuming classical strategies (i.e. RFLP typing) and the need for high quantities of DNA template.     

DHPLC-mtDNA控制区多态性分析系统的建立

目的基于变性高效液相色谱技术,建立一种不需测序和杂交的新的mtDNA控制区多态性分析系统。方法mtDNA控制区序列(包括HVⅠ,HVⅡ和HVⅢ)被分为4个扩增片段,采用配对分析突变检测模式进行DHPLC分析。对DHPLC检测条件(包括柱温和洗脱梯度等)进行优化。对100个不同类型差异序列的组合配对以检验该方法的检测效力。结果10组序列相同的样本配对DHPLC图谱均只显示1个样品峰。对序列相差1个碱基～7个碱基、插入(/缺失)1个碱基、插入(/缺失)2个碱基等类型的90个扩增片段组合,用DHPLC进行分析,均得到≥2个样本峰的DHPLC图谱,序列差异检出率达100%。该技术可检测的异质性DNA成分的最小百分含量为10%。结论DHPLC-mtDNA控制区多态性分析系统快速、经济和实用,在检测mtDNA异质性方面较直接测序更灵敏。     

中国汉族人mtDNA控制区异质性遗传规律

目的探讨中国汉族人mtDNA控制区异质性分布情况和遗传规律。方法将人mtDNA控制区扩增成6个部分互相重叠的片段,利用已建立的DHPLC技术分析其异质性规律。结果对150例汉族无关个体的多种组织检测,发现异质性个体的发生率达34%(51/150);个体的组织mtDNA异质性检出率最高为脑(50/150)、心肌(48/150)、最低为骨骼(22/150);本组共发现mtDNA控制区异质性位点有36个;同一个体可有多个异质性位点,最多的不超过3个;未发现异质性发生率有性别差异;超过41岁的高年龄组的异质性发生率(27/59)高于低年龄组(24/91);同一个体在2年前后取的血样,异质性检测结果一致;同一母系不同成员的异质性位点相同,但异质性mtDNA的含量有差异。结论DHPLC检测mtDNA控制区异质性具有高分辩力;mtDNA控制区异质性在中国汉族人中广泛存在;上述结果可作为mtDNA控制区多态性作个人认定和亲权鉴定的指导性资料。     

潍坊地区六种常见嗜尸性蝇类mtDNA COI区序列研究

目的解决蝇类成虫、蛹和蛆快速、准确鉴定的难题。方法随机采集潍坊地区人尸周围环境中的蝇类成虫、蛹和蛆,提取其mtDNA,经PCR扩增、产物检测与纯化、测序等,比较其序列差异。结果六种常见嗜尸性蝇类相互之间序列差异明显,同一种蝇的成虫、幼虫、蛹之间未见差异。结论mtDNACOI区序列分析可作为法医学嗜尸性蝇类种属鉴别的可靠方法。