7个Y-STR基因座荧光标记复合分型及在中国汉族群体中遗传多态性的研究

建立了FAM（蓝色荧光）标记的DYS456、DYS464a/b/c/d和DYS527a/b和JOE(绿色荧光)标记的DYS531、DYS709、DYS448和DYS522 7个Y染色体STR基因座(相当于11个位点)复合扩增分型体系。利用ABI3100遗传分析仪调查中国广东汉族151例和河南106例汉族无关男性个体的遗传多态性分布, 并探讨在法医学中的应用价值。结果显示, 此方法能作出正确分型的最低基因组DNA量为0.02 ng; 在女性成份为男性成份150倍的混合DNA样本(总DNA量为160 ng)中能正确检出全部7个Y-STR基因座的基因型。 广东和河南2个汉族男性群体中, 观察到的单体型分别有150、105种, 其中仅观察到1次的单体型分别有149、104种, 单体型多样性(HD)分别为0.999912、0.999820; 这组Y-STR单体型在两个群体中的分布差异有统计学显著意义(秩和检验: P<0.001)。此7个Y-STR基因座多态性分析适用于法医学实践和人类进化的研究。     

对法医学相关的DYS549、DYS527和DYS459基因座在男性不育症人群中的缺失、重复调查

位于Y染色体无精症因子区域(Azoospermia factor, AZF)的基因座位点DYS549、DYS527和DYS459在法医学鉴定和家系分析中被广泛应用。但是,在男性不育患者中,DYS549、DYS527和DYS459位点很可能会表现出特殊的基因型,对应用Y染色体短串联重复序列(Y chromosome short tandem repeat, Y-STR)进行个体识别的结果产生干扰。因此,文章应用14个Y-STR基因座复合扩增体系和Y染色体AZFc区DAZ、CDY1基因的拷贝数检测等方法,探讨男性不育症中法医学相关的3个Y-STR基因座的异常分型,对个体识别和家系分析中的DNA检验异常结果提供合理的解释。在240例男性非梗阻性无精、严重少精、先天性双侧输精管缺如(CBVAD)患者中,采用改良的多重PCR体系进行AZF区域微缺失的序列标签位点(Sequence tagged sites, STSs)检测,发现AZF微缺失40例(AZFa：2例;AZFb：2例;AZFc：30例;AZFb+c：6例),AZF的总缺失率为16.67%。应用14 Y-STR复合扩增体系对上述AZF微缺失的阳性患者样本进行检测,发现所有AZFb缺失患者存在DYS549等位基因缺失,AZFc缺失患者存在DYS527、DYS459等位基因缺失,AZFb+c缺失患者存在DYS549、DYS527和DYS459等位基因缺失。在AZF微缺失阴性的不育症患者中,通过检测DAZ、CDY1基因拷贝数发现10例AZFc部分复制的患者(1例为先天性输精管缺如,2例非梗阻性无精症,7例严重少精子症),占所调查不育人群的4.17%。男性不育人群AZF区域3个Y-STR基因座多态性会造成等位基因缺失或者重复,这些异常分型是由于临床遗传缺陷造成的而不是实验偏差。阐明Y-STR在男性不育人群中的异质性可以更好地完善Y-STR数据库和解释STR实验结果。     

广东汉族22个Y-STR基因座遗传多态性及遗传关系分析

调查了广东汉族群体22个 Y-STR基因座的遗传多态性分布情况, 探讨其群体遗传学及法医学应用价值。通过自行建立的两组Y-STR荧光标记复合扩增体系(MultiplexⅠ: DYS505, DYS533, DYS576, DYS588, DYS634, DYS643; MultiplexⅡ: DYS461, DYS481, DYS504, DYS508, DYS607)和应用进口Powerplex Y System (DYS19, DYS389Ⅰ/Ⅱ, DYS390, DYS391, DYS392, DYS393, DYS385, DYS437, DYS438, DYS439), 对广东汉族216 名无关男性个体进行22 个STR基因座的复合分型, 用ABI310基因分析仪对扩增产物进行检测, 统计22 个Y-STR基因座的群体遗传学参数, 并结合已公开发表的其他12 个群体“扩展单倍型”的数据资料, 分析广东汉族群体遗传距离和聚类关系。3 组复合扩增系统均可成功进行分型, 基因多样性GD值在0.3299(DYS634)~ 0.9425(DYS385); 22 个Y-STR基因座共同构成的单倍型214 种, 单倍型多样性为0.9999。广东汉族和潮汕汉族的遗传距离最近(-0.0030), 与东北汉族的遗传距离最远(0.0195)。22 个Y-STR基因座联合检测具有丰富的遗传多态性, 对建立Y染色体STR数据库, 研究群体遗传学和进行法医学应用有重要意义。     

北方汉族人群8个多拷贝Y-STR基因座的遗传多态性

为分析8个多拷贝Y-STR基因座在北方汉族群体中的遗传多态性和突变率,并同其他群体进行群体间差异分析,探讨其法医学和群体遗传学研究的应用价值。采用复合扩增体系对508名北方汉族无关男性个体和156对父子的8个多拷贝Y-STR基因座进行扩增,应用ABI3730分析仪进行基因分型,采用相应软件计算基因(单倍型)频率、基因多态性和突变率并对群体间差异进行分析;北方汉族群体的8个多拷贝Y-STR基因座的基因多态性普遍较高,基因多态性在0.673 8(DYS389I)~0.968 8(DYS464)之间,共检出484个单倍型,单倍型多态性为0.9998,家系调查中共发现5次基因突变,其中,各个基因座的每代突变率在0~9.62×10~(-3)(DYS535)之间,总平均突变率为1.60×10~(-3)(95%CI 0.5-3.7×10~(-3));群体间比较显示:8个基因座的遗传多态性在群体间存在差异,遗传距离在0.001 64~0.098 02之间。北方汉族群体的8个多拷贝Y-STR基因座的基因多态性普遍较高,适合应用于法医学个体识别和亲权鉴定中。     

福建畲族群体17个Y-STR基因座单倍型及遗传关系

应用Y-filerTM试剂盒及基因分型技术,检测152份福建畲族无关男性个体17个Y-STR基因座的多态性分布,计算等位基因频率及单倍型多样性,并结合已公开发表的其他11个群体相应基因座的单倍型资料,分析福建畲族群体遗传距离和聚类关系。福建畲族DYS385a/b基因座检出50种单倍型,其余15个Y-STR基因座分别检出3-11个等位基因,基因多样性GD值在0.4037(DYS391)～0.9725(DYS385a/b);观察到DYS19和DYS390基因座双等位基因和DYS385a/b基因座三等位基因,以及DYS448等部分基因座出现的"off-ladder"等位基因现象。17个Y-STR基因座共同构成的单倍型144种,其中138种单倍型出现1次,5种出现2次,1种出现4次,累计GD值为0.9990。从遗传距离分析发现,福建畲族与浙江汉族之间的遗传距离最近(0.0042),与青海藏族(0.2378)之间的遗传距离相对较远。福建畲族最靠近由台湾群体、浙江汉族、南方汉族等典型南方汉族群体聚成的分支区域。结果表明该17个Y-STR基因座在福建畲族群体中具有丰富的遗传多态性,对建立Y染色体STR数据库,研究群体遗传学和进行法医学应用有重要意义。     

大仓鼠DNA 指纹谱探针的筛选

采用一种简便提取高质量DNA 的方法, 从大仓鼠肝脏组织中提取其总DNA , 分别以人工合成的微卫星核心序列(GTG)₅和(CA)₈做单一引物, 进行特异引物PCR 反应。电泳检测后回收15 条特异性片段。与被标记过的大仓鼠基因组DNA 反向杂交结果表明, 15 个片段中(GTG)_5-8 、(CA)_8-1b和(CA_{) 8-5b}产生了较强的阳性信号。我们依据3 个片段的测序结果设计适合DIG标记的探针, 该探针得到的大仓鼠不同地理种群个体的指纹图谱有较高的个体特异性和种群多态性, 而且与传统的来源于其它生物重复序列的探针如33.6 和33.15 形成的指纹图谱相比得到的变异适中, 便于统计。     

中国云南白族和新疆维族7个Y-STR基因座的遗传多态性

利用AB I3100遗传分析仪, 我们调查了7个Y-STR基因座构成的单体型在中国云南白族133例和新疆维族105例无关男性个体中的分布。7个Y-STR基因座包括DYS456、DYS464 a/b/c/d、DYS527 a/b、DYS531、DYS709、DYS448和DYS522。结果显示,云南白族和新疆维族2个民族男性群体中,分别观察到133、105种单体型, 均为仅观察到1次的单体型,单体型多样性(HD)均趋近于1, 比较这种单体型在两个群体中的分布差异,显示有统计学显著意义(秩和检验: P<0.001)。中国云南白族和新疆维族7个Y-STR基因座具有较高的多态性, 为法医学实践和群体遗传学的研究提供了有用的基础资料。     

广州汉族人群D12S391和D11S554基因座序列变异

为研究高度变异的sTR基因座核心序列结构,对广州汉族人群突变率较高的D12S391和D11S554基因座等位基因进行了序列分析。结果显示D12S391基因座核心序列结构为(AGAT)8～17(AGAC)6～10(AGAT)0～1,其较小片段等位基因(15～18)仅表现为第1个重复单位(AGAT)数目的变异,而较大片段等位基因(19～27),可表现为第2或第3个重复单位数目的变异。发现有4种新的等位基因,分别命名为22″、23″、24′″和27。D11S554基因座核心序列结构更复杂,有5种核心序列,其中3种具有相同的基本结构(AAAGG)(AAAG)4(AAAGG)2～3,(AAAG)13～19。其大片段等位基因(219～249)核心序列结构中,既有四核苷酸重复,还有五核苷酸重复,以及单个硷基的变异、硷基插入或缺失。两基因座均存在序列异质性。结果表明D12S391和D11S554基因座属复杂重复类型,为其准确分型增加了难度,首先需建立相应群体的等位基因分型标准物。     

山西汉族17个Y-STR基因座遗传多态性及遗传关系

为了调查山西汉族群体17个Y-STR基因座的多态性分布,探讨其群体遗传学及法医学应用价值,文章应用Y-filer TM试剂盒检测222名山西汉族无关男性个体的17个Y-STR基因座,用ABI3130遗传分析仪进行基因分型,计算等位基因频率及单倍型多样性,并结合已公开发表的国内其他13个群体相关数据资料,分析山西汉族群体遗传距离和聚类关系。结果:山西汉族个体中共检出219种单倍型,单倍型多样性为0.9999;基因多样性GD值在0.3894(DYS391)~0.9755(DYS385a/b)。从遗传距离分析发现,山西汉族与吉黑汉族之间的遗传距离最近(?0.0001),与台湾群体(0.0152)之间的遗传距离相对较远。结果表明该17个Y-STR基因座在山西汉族群体中具有丰富的遗传多态性,对建立Y染色体STR数据库、研究群体遗传学和进行法医学应用有重要意义。     

云南泸西汉族17个Y-STR基因座多态性及遗传关系分析

调查云南泸西县汉族群体17个Y-STR基因座的多态性分布, 探讨其群体遗传学及法医学应用价值。应用AmpF?STR?Yfiler荧光标记复合扩增系统扩增156名汉族无关男性个体的17个Y-STR基因座, 用ABI3100遗传分析仪进行基因检测, 计算等位基因及单倍型频率, 并结合已公开发表的国内外其他16个群体的遗传学资料, 分析各群体间的遗传距离。云南泸西汉族男性群体中, 共观察到154种单倍型, 其中152种为仅观察到1次的单倍型, 2种单倍型观察到2次, 单倍型多样性(HD)值为0.9998, 基因多样性(GD)值在0.3901(DYS437)—0.9632(DYS385a/b)。17个群体遗传距离分析提示, 国内人群云南泸西汉族与湖南汉族的遗传距离最小(0.005), 与闽南汉族的遗传距离最大(0.035); 在中国周边群体中,云南泸西汉族与新加坡华人的遗传距离最小(0.015), 与日本人和马来西亚印度人群的遗传距离最大(0.060)。结果表明, 17个Y-STR基因座在云南泸西汉族中具有较高的遗传多态性, 适用当地的法医学应用。与其他民族群体的遗传多样性比较, 对了解各群体的起源、迁徙及相互关系有重要意义。     

DYS19 and DYS199 loci in a Chilean population of mixed ancestry

The current Chilean population originated from admixture between aboriginal populations (Amerindians) and Spanish conquerors of European origin. Consequently, the unions that gave rise to the Chilean population were chiefly between Spanish males and aboriginal females, and not the converse. To test the hypothesis that the Y chromosome of the Chilean population is mainly of Spanish origin, while the other chromosomes are from mixed (European and aboriginal) origin, we studied the DYS19 and DYS199 loci in two samples. One sample was obtained from a high socioeconomic stratum, while a second sample was from a low stratum. We studied male blood donors (N = 187) from Santiago, the capital of the country. Subjects were typed for the autosomal ABO and Rh (locus D) blood groups, and for the Y-linked DYS19 and the DYS199 loci, reported as Y-chromosome haplotypes. The aboriginal admixture was estimated for each genetic marker. The percentage of aboriginal admixture was 38.17% for the ABO system and 31.28% for the Rh system in the low socioeconomic stratum and 19.22% and 22.5%, respectively, in the high stratum. Y-chromosome haplotype frequencies constructed from the DYS19 and DYS199 loci demonstrated that the main haplotypes were DYS19*14/DYS199 C, as is often the case with many European populations, and DYS19*13/DYS199 C. The aboriginal admixture from Y-haplotype frequencies was estimated to be 15.83% in the low socioeconomic stratum and 6.91% in the high stratum. These values are lower than the values found using autosomal genetic markers, and are consistent with the historical background of the population studied. This study highlights the population genetic consequences of the asymmetric pattern of genome admixture between two ancestral populations (European and Amerindian).     

Highly polymorphic STR marker amplified with human DYS389 primers in Japanese macaques (Macaca fuscata)

Amplification products from male and female Japanese macaques were obtained by PCR with human Y-chromosomal DYS389 primers. These products were examined by electrophoresis and sequence analysis. The PCR products from the 12 Japanese macaques tested had different band patterns on an electrophoretogram. Sequence analysis of the products revealed that the high polymorphism originated from variable numbers of repeats of two separate CTAT sequences. The sequences of the Japanese macaque products were similar to those of the reference human DYS389 sequence. However, variable CTGT repeats and a difference in the second forward primer binding site yielded two products in human males, DYS389I and DYS389II, which do not exist in Japanese macaques. Our results suggest that the human DYS389 primers may be a potential tool not only for distinguishing between human and Japanese macaque DNA samples, but also for identifying individual macaques, because of the highly polymorphic alleles.     

陕西渭南地区汉族人群17个Y-STR基因座多态性及遗传关系分析

调查陕西渭南地区汉族群体17个Y-STR基因座的多态性,探讨其群体遗传学及法医学应用价值。应用Y-fi ler荧光标记复合扩增系统,对413名陕西渭南地区汉族无关男性个体17个Y-STR基因座进行复合扩增,用ABI3130遗传分析仪进行基因分型,计算各基因座的群体遗传学参数,并结合已经发表的其他10个群体相应基因座的单倍型资料,分析各群体间的遗传距离。413名陕西渭南汉族个体共检出405种单倍型,其中397种单倍型仅出现1次,单倍型多样性达0.9999,基因多样性(GD)为0.4130(DYS391)~0.9734(DYS385a/b),累计GD值为0.9999。遗传距离分析提示,陕西渭南汉族与辽宁满族的遗传距离最小(0.00110),与青海藏族的遗传距离最大(0.22333)。结果表明,17个Y-STR基因座在陕西渭南汉族群体中具有丰富的遗传多态性和较高的非父排除能力,在法医学和人类群体遗传学研究中具有重要价值。     

Molecular characterization and population study of an X chromosome homolog of the Y-linked microsatellite DYS391

A novel microsatellite homologous to DYS391, a (GATA)(n) short tandem repeat on the human Y chromosome, was identified and characterized in the present work. Employing somatic cell hybrid and deletion panels in a PCR-based approach, we found out that the new microsatellite is located in Xp21.2-22.3, while its Y counterpart mapped to Yq11.21. This X-linked locus (provisionally called DXYS391) and its Y homolog constitute one more example of similarity outside the pseudoautosomal regions between the two human sex chromosomes. Sequencing data showed high levels of homology in the flanking regions of DXYS391 and DYS391 that differ primarily by the presence of a (GACA)(3) motif in the Y locus. Both loci were detected in chimpanzee DNA, suggesting that a putative transposition from the X to the Y occurred before the human/chimpanzee split. The allele frequencies of DYS391 and DXYS391 were investigated, respectively, in 271 Y and 337 X chromosomes from distinct human populations worldwide. DYS391 consistently displayed greater among-population component of the variance of the allele frequencies than DXYS391, as expected due to the three-times lower effective population size of Y chromosomes relative to the X. The intra-population diversity of DYS391, measured by Nei's locus diversity as well as by allele size variance, was lowest in Amerindians, while very low diversity of DXYS391 was seen in Africans. Since our African data are based on a small sample, further studies will be necessary to evaluate better this observation.     

贵州三都水族Y染色体上七个STR基因座的遗传多态性分析

采用多重PCR技术, 结合ABI PRISMTM 377 DNA Sequencer四色荧光标记进行基因扫描分型, 对中国贵州三都水族群体进行7个Y-STR基因座的多态性分析, 计算其基因频率、遗传多样性及单倍型多样性, 获取相应的遗传多态信息。结果显示: 在94个无关男性样本中, DYS19、DYS389Ⅰ、DYS389Ⅱ、DYS390、DYS391、DYS392、DYS393等基因座分别检出6, 4, 6, 2, 3, 5, 4种等位基因, 遗传多样性在0.124(DYS389Ⅰ)~0.630(DYS19)之间; 由此组成的单倍型为27种, 单倍型多样性0.868。此7个Y-STR基因座在贵州三都水族群体中具有较好的多态性, 单倍型具有较高的遗传多态。     

Phylogenetic resolution of complex mutational features at Y-STR DYS390 in aboriginal Australians and Papuans

Y-chromosomal short tandem repeats (STRs) are used for the study of male aspects of human evolution as well as for forensic applications and paternity testing. Both applications require an understanding of the underlying mutational mechanisms that create variability. We describe complex mutations at the substructured DYS390 STR locus in 97 natives of the New Guinea/Australian region. Sequencing of short alleles in these populations indicates multirepeat deletions. All samples are further characterized using the five additional Y-STR loci DYS19, DXYS156-Y, DYS391, DYS392, and DYS393. Phylogenetic analysis of the resulting haplotypes yields ethnically specific clusters predating the settlement of Australia and Papua New Guinea (although archaic Homo sapiens or Homo erectus lineages are absent). The phylogeny confirms that DYS390 violates the stepwise mutation model and demonstrates that the DYS390 locus mutates relatively rapidly and retains its variability after structural change.