Y-STR基因座多态性分析及在法医学上的应用研究

发布时间：2018-03-29 00:24

本文选题：Y染色体　切入点：Y-STR　出处：《山西医科大学》2009年硕士论文

【摘要】： 目的Y染色体遗传标记多态性较低,需要寻找更多的基因座用于遗传学分析。本实验从染色体基因座中筛选出3个新Y-STR基因座,调查这3个Y-STR基因座在山西汉族人群中的等位基因频率及单倍型分布,并对这3个Y-STR新基因座在法医学上的应用进行可行性探讨。方法(1)样本采集:随机抽取120例山西汉族健康男性无关个体静脉血进行等位基因频率及单倍型调查;随机抽取20例山西健康女性血样进行阴性对照;采集20例两代家系血样进行突变观察;采集同一例健康男性尸体的血液和组织器官进行同一性观察;采集3例常见雄性动物血液进行种属特异性观察。(2)用有机溶剂提取法提取基因组DNA;根据GDB基因库提供的引物序列和自行设计的引物序列合成引物。(3) PCR扩增后,进行扩增产物的非变性聚丙烯酰胺凝胶(PAGE)电泳分离和硝酸银染色分型,测序。(4)计算其法医学参数。结果研究发现DYS708基因座在120例山西汉族男性群体中分别检测出6个等位基因, DYS708*24、DYS708*25、DYS708*26、DYS708*27、DYS708*28、DYS708*29,其等位基因频率见表1。在山西汉族群体中,DYS708基因座的基因多样性(GD)为0.823,其个人识别能力(DP)和非父排除率(PE)为0.823; 20例女性对照无特异性扩增产物且20例两代家系调查无突变。同一男性个体血液、组织器官检测结果分型一致。3例动物血样中均未观察到特异性扩增。DYS724,32例样本观察到1个等位基因,68例样本观察到2个等位基因, 20例样本观察到3个等位基因,测序无结果。DYS725扩增目的DNA在聚丙烯凝胶上出现两条带,且所有样本无基因多态性。结论基因座DYS708具有较高的个人识别能力,所得到的数据可为山西汉族人群法医学个人识别、亲子鉴定及遗传学研究提供依据。DYS724,DYS725基因多态性低,不宜作为法医学个人识别、亲子鉴定及遗传学研究的依据。目的在前期研究的基础上筛选出12个遗传多态性较高的Y-STR基因座,调查这12个Y-STR基因座在山西汉族人群中的等位基因频率及单倍型分布,并对这12个基因座单倍型在法医学上的应用进行可行性探讨。方法(1)样本采集:随机抽取120例山西汉族健康男性无关个体静脉血进行等位基因频率及单倍型调查;随机抽取20例山西健康女性血样进行阴性对照;采集20例两代家系血样进行突变观察;采集同一例健康男性尸体的血液和组织器官进行同一性观察;采集3例常见雄性动物血液进行种属特异性观察。(2)用有机溶剂提取法提取基因组DNA;根据GDB基因库提供的引物序列和自行设计的引物序列合成引物。(3) PCR扩增后,进行扩增产物的非变性聚丙烯酰胺凝胶(PAGE)电泳分离和硝酸银染色分型,测序。(4)计算其法医学参数。结果研究发现这12个基因座(DYS650, DYS651,DYS708,DYS711,DYS713, DYS714,DYS715,DYS716,DYS717,DYS718,DYS721,DYS726)在120例山西汉族男性群体中分别检测出4、5、6、6、8、6、6、5、5、5、4、5、5、4个等位基因,它们分别是DYS650*14、DYS650*15、DYS650*16、DYS650*17;DYS651*7、DYS651*8、DYS651*9、和DYS651*10、DYS651*11; DYS708*24、DYS708*25、DYS708*26、DYS708*27、DYS708*28、DYS708*29;DYS711*60、DYS711*63、DYS711*65、DYS711*68、DYS711*70、DYS711*74;DYS713*36、DYS713*38、DYS713*40、DYS713*42、DYS713*44、DYS713*46;DYS714*10、DYS714*11、DYS714*12、DYS714*13、DYS714*14、DYS714*15;DYS715*12、DYS715*13、DYS715*14、DYS715*15、DYS715*16;DYS716*15、DYS716*16、DYS716*17、DYS716*18、DYS716*19;DYS717*18、DYS717*19、DYS717*20、DYS717*21、DYS717*22;DYS718*14、DYS718*15、DYS718*16、DYS718*17; DYS721*9、DYS721*10、DYS721*11、DYS721*12和DYS721*13;DYS723*10、DYS723*11、DYS723*11、和DYS723*13、DYS723*14; DYS726*11、DYS726*12、DYS726*13、DYS726*14;其等位基因频率见表14。在山西汉族群体中,Y-STR基因座的基因多样性(GD)最低为0.730(DYS650) ,最高为0.831 (DYS711),其个人识别能力(DP)和非父排除率(PE)最低为0.730(DYS650) ,最高为0.831 (DYS711);12个Y-STR基因座共同构成的单倍型为120种,其多样性(HD)为0.99。20例女性对照无特异性扩增产物且20组两代家系调查无突变。同一男性个体血液、组织器官检测结果分型一致。3例动物血样中均未观察到特异性扩增。结论这12个Y-STR基因座单倍型具有较高的个人识别能力,所得到的数据可为山西汉族人群法医学个人识别、亲子鉴定及遗传学研究提供依据。
[Abstract]:The purpose of the Y chromosome genetic polymorphism is low, need to find more loci for genetic analysis. The results from chromosomal loci screened 3 Y-STR loci, this survey of 3 Y-STR loci in Shanxi Han population allele frequencies and haplotype distribution, and Discussion on these 3 Y-STR new loci in forensic application of feasibility.
Methods (1) specimens were collected from 120 cases of healthy male unrelated individuals in Shanxi Han blood allele frequencies and haplotype survey; randomly selected 20 cases of healthy women in Shanxi blood samples were collected from 20 patients with negative control; the two generation family of blood samples were collected from the same mutation observed; healthy male body blood and organs observe the same; collecting 3 cases of common male animal blood species specific observation. (2) genomic DNA was extracted by organic solvent extraction; primers according to the sequence of GDB gene library and self-designed primer sequence primers. (3) after PCR amplification, non denaturing polyacrylamide gel products the (PAGE) electrophoresis and silver nitrate staining, DNA sequencing. (4) calculate the forensic parameters.
The study found that DYS708 loci in 120 cases of Shanxi Han male population were detected 6 alleles, DYS708*24, DYS708*25, DYS708*26, DYS708*27, DYS708*28, DYS708*29, the allele frequencies are shown in table 1. in Shanxi Han population, DYS708 locus gene diversity (GD) was 0.823, the personal identification ability (DP) and the probability of exclusion (PE) was 0.823; 20 cases of female were no specific amplification products and 20 cases of the two generation pedigree investigation without mutation. The same male individual blood, tissue typing results.3 cases in animal blood were not observed in the specific amplification of.DYS724,32 cases samples of 1 alleles were observed in 68 samples, 2 alleles were observed in 20 samples, 3 alleles were observed, no sequencing results of.DYS725 amplification DNA two bands in polyacrylamide gel, and all samples without gene polymorphism.
Personal identification conclusion locus DYS708 is higher, the data obtained can be personal identification for forensic in Shanxi Han population, provide the basis for the study of.DYS724 paternity and genetic polymorphism of DYS725 gene is low, not as a forensic personal identification, paternity studies and genetic basis.
Objective on the basis of previous research selected Y-STR loci of 12 higher genetic polymorphism, this survey of 12 Y-STR loci in Shanxi Han population allele frequencies and haplotype distribution, and study of the 12 haplotypes of feasibility in forensic application.
Methods (1) specimens were collected from 120 cases of healthy male unrelated individuals in Shanxi Han blood allele frequencies and haplotype survey; randomly selected 20 cases of healthy women in Shanxi blood samples were collected from 20 patients with negative control; the two generation family of blood samples were collected from the same mutation observed; healthy male body blood and organs observe the same; collecting 3 cases of common male animal blood species specific observation. (2) genomic DNA was extracted by organic solvent extraction; primers according to the sequence of GDB gene library and self-designed primer sequence primers. (3) after PCR amplification, non denaturing polyacrylamide gel products the (PAGE) electrophoresis and silver nitrate staining, DNA sequencing. (4) calculate the forensic parameters.
The results of the study found that the 12 loci (DYS650, DYS651, DYS708, DYS711, DYS713, DYS714, DYS715, DYS716, DYS717, DYS718, DYS721, DYS726) in 120 cases of Shanxi Han nationality male group were detected 4,5,6,6,8,6,6,5,5,5,4,5,5,4 alleles, which respectively is DYS650*14, DYS650*15, DYS650*16, DYS651*7, DYS650*17; DYS651*8, DYS651*9, and DYS651*10, DYS651*11; DYS708*24, DYS708*25, DYS708*26, DYS708*27, DYS708*28, DYS708*29; DYS711*60, DYS711*63, DYS711*65, DYS711*68, DYS711*70, DYS711*, DYS713*36, DYS713*38, 74; DYS713*40, DYS713*42, DYS713*44, DYS713*46; DYS714*10, DYS714*11, DYS714*12, DYS714*13, DYS714*14, DYS714*15; DYS715*12, DYS715*13, DYS715*14. DYS715*15, DYS715*16; DYS716*15, DYS716*16, DYS716*17, DYS716*18, DYS716*19; DYS717*18, DYS717*19, DYS717*20, DYS717*21, DYS717*22; DYS718*14, DYS718*15, DYS718*16, DYS718* 17; DYS721*9, DYS721*10, DYS721*11, DYS721*12 and DYS721*13; DYS723*10, DYS723*11, DYS723*11, and DYS723*13, DYS723*14; DYS726*11, DYS726*12, DYS726*13, DYS726*14; the allele frequencies are shown in table 14. in Shanxi Han population, Y-STR locus gene diversity (GD) for a minimum of 0.730 (DYS650), the highest 0.831 (DYS711), the personal identification ability (DP) and the probability of exclusion (PE) for a minimum of 0.730 (DYS650), the highest was 0.831 (DYS711); the 12 Y-STR loci haplotypes into 120 types, the diversity (HD) for 0.99.20 patients and 20 female control survey result two generation family specific amplification without mutation. The same male individual blood, tissue typing results.3 cases in animal blood were observed in specific amplification.
Conclusion these 12 Y-STR loci haplotypes have high ability of personal identification. The data obtained provide evidence for forensic identification, paternity testing and genetic research in Shanxi Han population.

【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919

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