【作者】 王文菲；

【导师】 曾昭书；

【作者基本信息】 郑州大学 ， 法医学， 2012， 硕士

【摘要】 背景和目的短串联重复序列一般是由两个或多个核苷酸为核心序列的重复出现而组成的串联结构,以重复区域为中心的片段长度一般在400bp以内,重复单位为2～6bp,重复次数可达10-60次。由于不同STRs位点的核心序列不同,而同一STR位点在不同人中的重复次数不同,导致出现个体差异,构成了长度多态性,广泛应用于个体识别和亲权鉴定。由于STRs具有分布广、信息量大、高度多态性以及遵循孟德尔共显性遗传等特点,也成为推断民族遗传距离的有效工具。哈萨克族是哈萨克斯坦的主要民族和中国的少数民族,人口1700万。在中国主要分布于新疆维吾尔自治区伊犁哈萨克自治州、木垒哈萨克自治县、巴里坤哈萨克自治县、甘肃省阿克塞哈萨克族自治县。该民族使用文字是以阿拉伯字母为基础的哈萨克语,按语言系属分类,属于突厥语族人种。按血统分类,属于蒙古人种北亚类型和欧罗巴人种及印度地中海类型之间的混合类型,属于混血民族。由于部分哈萨克人仍以游牧生活为主,无固定居住区域,且地理位置较为偏僻,交通不便,经济发展欠发达等多重原因,我国哈萨克族人群的15个基因座数据资料至2011年10月一直未见报道。因导师组有机会从新疆合作单位取得新疆哈萨克族血样,特对这些血样进行法医学常用STR基因座的检测。然后,本研究通过公用数据库或同行评议期刊等网络资料库,收集了中国各个省份、其它各个民族的STR基因座频率数据。通过对40个不同民族群体间遗传距离的计算,构建了多个民族间的系统发生树,以探讨其遗传关系。方法使用酚-氯仿法对100例新疆哈萨克族外周血样进行DNA提取,使用Identifiler试剂盒进行荧光标记PCR扩增,而后使用3130DNA测序仪对PCR产物进行毛细管电泳,最后完成对新疆哈萨克族100例健康无亲缘性个体的15个STR基因座遗传多态性的研究。通过公用数据库或同行评议期刊等资料库收集中国各个省份、各个民族其它群体的STR基因座频率数据；将获得的文献数据与新疆哈萨克族的遗传数据,筛选汇总后计算出9个法医学常用STR位点(D13S317.D18S51.D21S11.D3S1358. D5S818.D7S820.D8S1179.FGA.vWA)在中国40个民族中的基因频率,以phyl ip3.69统计软件包计算遗传距离,并通过Neighbor－joining法构建系统发生树,研究各民族间的遗传关系。结果对100例新疆哈萨克族的15个STR基因座进行检测分别检出了10、15、8、10、6、6、7、6、12、11、8、6、12、7、10个等位基因,其基因频率分布范围分别在0.01-0.25,0.005-0.275,0.01-0.225,0.015-0.33,0.01-0.335,0.01-0.285,0.015-0.24,0.025-0.265,0.015-0.205,0.015-0.29,0.02-0.26,0.01-0.595,0.005-0.255,0.025-0.365以及0.005-0.205之间.比较所有40个族群的9个STRs基因座基因频率数据计算出了遗传距离,并根据遗传距离采用Neighbor-joining法构建了40个族群之间的系统发生树。在树状图上,可以看出在系统发生树的下半部分,新疆维吾尔族先与内蒙古蒙古族首先聚合,然后再与新疆哈萨克族、内蒙古达斡尔族、天津汉族,青海回族分别聚合,继而与青海藏族、西藏藏族、西藏珞巴族,浙江畲族的亚类相聚合,再与陕西汉族、云南白族,天津朝鲜族、湖南土家族、山东汉族、江苏汉族相聚合。然后与河北汉族、内蒙古汉族、河南汉族,辽宁汉族形成的汉族亚类相聚合,构成了整个北方人群。系统发生树上半部分,广西毛南族和广西京族首先聚合,继而与广西苗族、广西仫佬族聚合,与广西瑶族和海南黎族聚合的亚类聚合。接下来与广西瑶族、海南汉族相聚合。广西侗族、广西壮族。广西汉族、广东汉族的亚类与之相聚合后,又与四川汉族、福建汉族以及湖北汉族以及重庆土家族聚合,构成南方人群。而安徽汉族、浙江汉族,构成了南方汉族和北方汉族之间的过渡人群。结论通过对100例新疆哈萨克民族人群血样15个STR基因座数据资料的检测,可以为法医学个体识别、亲权鉴定、DNA数据库技术等法医学应用提供基础资料,同时也为研究该民族基因特异性,分析人类进化、民族起源、族群之间的亲缘关系以及由于民族基因缺陷而特有的疾病谱提供了重要依据。中国南北人群之间存在遗传差异,汉族和少数民族之间存在遗传差异更多还原

【Abstract】 Background and purposeShort Tandem Repeats (STR) is a kind of DNA sequences that is generally formed by the repeat of a core sequence with two or more nucleotides. The fragment length of the repeat region is generally less than400bp, the core repeat unit is2～6bp and repetitions can be10～60times. Different STRs have different core sequences, and the same STRs have different repetitions among persons. These lead to the individual differences and form a genetic polymorphism. So STRs are widely used in individual identification and paternity testing. Because STRs have a wide distribution, high polymorphism, Mendelian inheritance style, they have also become an effective tool to infer the genetic distance among different ethnic groups.Kazakh is Kazakhstan’s main ethnic group, and it is one of the minorities in China. The population of Chinese Kazakh is about17million. In China, this nationality mainly line in Xinjiang Uygur Autonomous Region, the Hi Kazakh Autonomous Prefecture, Mori Kazak Autonomous County, Barkol Kazak Autonomous County and Aksay Kazak Autonomous County in Gansu Province. Their native language is Kazakh, and the character of Kazakh this is based on Arabic alphabet. According to their inheritance linkage, this ethnic group belongs to a mixed type, which is among the North Asian type of the Mongoloid, the white race and the India Mediterranean race. It belongs to the Turkic tribe, classified by the linguisticians.Some Kazakh peoples is still living a nomadic way of life, with no fixed residential areas. Their areas are far more remote, with inconvenient transportation and less developed economic status. Due to these reasons, the15frequently used forensic STR data of the Kazakh population in China is not reported until we set out to test their samples at the end of2010. Since my supervisor have the opportunity to obtain Xinjiang Kazak blood smaples from his Xinjiang connections, we decide to test these blood samples for the data of the commonly used15forensic STR loci.Searching through the network databases or peer-reviewed journals, this study has collected the STR loci frequency data in other ethnic groups from all provinces in China. To explore the genetic relationship between the different Chinese nationalities, a phylogenetic tree based on the calculation of genetic distance among40different ethnic groups will be tried to be built.MethodsDNA samples were extracted from the peripheral blood of100Xinjiang Kazakh by the method of phenol-chloroform extraction. Using the Identifier kit to finish the fluorescence-PCR amplification, then a capillary electrophoresis for PCR products were done on3130DNA genetic analyzer. Finally, we accomplished the polymorphism investigation of the15STR loci among100healthy cases of Kazakh. Through the public databases or peer-reviewed journals, STR loci frequency data of other ethnic groups was collected from all provinces in China. After gathered the data from this study and references, we calculated the frequencies of9commonly used forensic STR loci (D13S317, D18S51, D21S11, D3S1358, D5S818, D7S820, D8S1179, FGA, vWA) in China’s40ethnic groups. The genetic distance was calculated by the Phylip3.69statistical package, and the phylogenetic trees was constructed by the Neighbor-joining method to study the genetic relationships between the various ethnic groups.Results15STR loci in100cases of Xinjiang Kazakh were detected, respectively, there are10,15,8,10,6,6,7,6,12,11,8,6,12,7,10alleles were found; the range of these gene frequencies distribute in0.01～0.25,0.005～0.275,0.01～0.225,0.015～0.33,0.01～0.335,0.01～0.285,0.015～0.24,0.025-0.265,0.015～0.205,0.015～0.29,0.02～0.26,0.01～0.595,0.005～0.255,0.025～0.365and0.005～0.205.Nine STRs loci gene frequency data of all40ethnic groups were compared to calculate the genetic distance, and a phylogenetic tree was constructed using the Neighbor-joining method based on genetic distances of40populations. In the lower half of the phylogenetic tree, the Xinjiang Uygur is the nearest to Mongolians, and then come near to the Kazak, to Inner Mongolia, to Daur, to Hans in Tianjin, to Hui in Qinghai, and then they go to Tibetans in Qinghai, Tibetans in Tibet, to Lhoba in Tibet, to Shes in Zhejiang subcategories for a convergence, then Shaanxi Hans, Yunnan Bais, Tianjin Koreans, Hunan Tujias, Shandong Hans, Jiangsu Hans. And then join Hans in Hebei, Hans in Inner Mongolia and Hans in Henan, Hans in Liaoning, constituting the entire population of North Chinese. On the upper half of the phylogenetic tree, the Maonan of Guangxi and the Jing of Guangxi come together first, the join the Guangxi Miao, Guangxi Mulao; then the Yaos in Guangxi and Lis in Hainan come in for a convergence. Next come the Yaos in Guangxi, Hans in Hainan. The Dongs in Guangxi, Zhuangs in Guangxi. Then Hans in Guangxi, Hans in Guangdong join, together with the Hans in Sichuan, Hans in Fujian, Hans in Hubei and Tujias in Chongqing, constituting the southern Chinese population. While Anhui Hans and Zhejiang Hans become a transition between the Southern Han Chinese population and Northern Hans.Conclusion The data of15STR loci in100cases of Kazakh obtained in this study will not only provide basic information for future’s forensic applications such as individual identification, paternity testing, but also provide an important basis for the study of this ethnic’s genetic characteristics, the analysis on its evolution and its origin, and the genetic relationships among different ethnic groups,and unique disease spectrum caused by ethnic genetic defects.There are genetic differences between the Northern and the Southern Chinese population.There are genetic differences between the Han population and the minorities.更多还原