1144 HEREDITAS (Beijing) allele and genotype frequency of nine X-STR loci among 120 unrelated individuals (55 female, 65 male) from Yugur ethn

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1 HEREDITAS (Beijing) , 30(9): ISSN 研究报告 DOI: /SP.J X-STR 陈艳炯 1, 陈峰 2, 辛娜 3, 张洪波 2, 郑海波 2, 余兵 2, 李生斌 2, 陈腾 1., ; 2., ; 3., : 为研究中国甘肃裕固族人群 X 染色体 STR 基因座的遗传多态性及其在群体遗传学中的应用, 采用 PCR 扩增, 变性聚丙烯酰胺凝胶电泳结合银染显带技术, 检测 120 名 ( 女 55, 男 65) 裕固族无关个体 9 个 X-STR 基因座 (DXS7130 DXS7132 DXS6804 DXS7423 DXS7424 DXS6789 DXS6799 DXS8378 和 HPRTB) 的等位基因频率及基因型分布, 以及存在连锁的 X-STR 基因座的单体型多态性 ; 同时, 利用 X-STR 构建系统发生树和进行聚类分析, 分析裕固族与我国其他民族的群体遗传关系结果发现, DXS7130 DXS7132 DXS6804 DXS7423 DXS7424 DXS6789 DXS6799 DXS8378 和 HPRTB 基因座分别检出个等位基因和种基因型, 9 个 X-STR 基因座女性的基因型频率分布均符合 Hardy-Weinberg 平衡 (P>0.05) 由 DXS7130 和 DXS8378 基因座组成的单体型共检出 15 种, 由 DXS6789 DXS6799 DXS7424 和 DXS6804 基因座组成的单体型共检出 55 种, 单体型多样性分别为和群体遗传多态性指标显示上述 9 个 X-STR 基因座均具有较高多态性, 在法医学个体识别亲权鉴定及群体遗传学研究中有重要应用价值对裕固族与我国其他民族群体遗传关系的研究结果显示, 裕固族与蒙古族及同处西北的汉族藏族关系较近, 而与回族维族关系较远, 提示裕固族是一个在起源上与蒙古族汉族以及藏族关系密切的民族群体 : X 染色体 ; 短串联重复序列 ; 单体型 ; 遗传多态性 ; 聚类分析 ; 中国裕固族 Genetic polymorphisms of X-STR loci in Chinese Yugur ethnic group and its application CHEN Yan-Jiong 1, CHEN Feng 2, XIN Na 3, ZHANG Hong-Bo 2, ZHENG Hai-Bo 2, YU Bing 2, LI Sheng-Bin 2, CHEN Teng 2 1. Department of Immunology and Pathogenic Biology, Xi an Jiaotong University School of Medicine, Xi an , China; 2. Department of Forensic Medicine, Xi an Jiaotong University School of Medicine, Xi an , China; 3. Clinical Laboratory, Affiliated Hospital of Xi an Medical College, Xi an , China Abstract: To study the genetic polymorphism of nine short tandem repeats (STRs) loci (DXS7130, DXS7132, DXS6804, DXS7423, DXS7424, DXS6789, DXS6799, DXS8378, and HPRTB) on X chromosome in Chinese Yugur ethnic group. The 收稿日期 : ; 修回日期 : 基金项目 : 教育部科技基础资源数据平台建设项目中华民族群体遗传资源数据整合共享平台 ( 编号 : ) 和陕西省科技攻关项目 ( 编号 : 2004K09-G12)[Supported by the Platform Construction Projects of Chinese Education Ministry for Sciences and Technological Fundamental Resources Data: Integrated and Participatory Data Platform for Chinese Population Genetic Resources ( ) and Shaanxi Key Project of Science and Technology ( 2004K09-G12)] 作者简介 : (1967 ),,,,, Tel: ; 通讯作者 : (1965 ),,,, Tel: ; chenteng@mail.xjtu.edu.cn

2 1144 HEREDITAS (Beijing) allele and genotype frequency of nine X-STR loci among 120 unrelated individuals (55 female, 65 male) from Yugur ethnic group were analyzed using PCR and followed by polyacrylamide gel electrophoresis and silver staining. The numbers of alleles in the nine X-STR loci were 8, 6, 6, 5, 6, 7, 6, 4, and 6, respectively; the numbers of genotypes in the nine loci were 16, 14, 13, 6, 13, 20, 11, 6, and 12, respectively. The genotype frequencies in females were in accordance with Hardy-Weinberg equilibrium (P>0.05). The nine X-STR loci were relatively abundant in polymorphic information for individual identification, paternity testing and population genetics. A total of 15 haplotypes were detected in DXS7130 and DXS8378 loci, and 55 haplotypes were detected in DXS6789, DXS6799, DXS7424, and DXS6804 loci. The haplotype diversity reached and , respectively. Phylogeny tree and cluster analysis based on X-STR allele frequencies in genesis showed that Yugur ethnic group share a close relationship with Mongolian ethnic group and Chinese Han, Tibetan population and far from Hui and Uygur ethnic group, who dwell in the northwest of China. Keywords: X-chromosome; short tandem repeats; haplotype; genetic polymorphism; cluster analysis; Chinese Yugur ethnic group,,,, (The international HapMap project, HapMap) [1] 3, DNA,,,,,, 44,,,,,, 9 STR [2], X 9 STR,,, DNA 120 ( 55, 65),, 3,, 2 ml, EDTA, DNA Chelex-100 [3] DNA 1.3 PCR PCR 12 μl, 1 PCR buffer(50 mmol/l KCl, 10 mmol/l Tris-HCl, ph 9.0), 1.5 mmol/l MgCl 2, 0.2 mmol/l dntp, 0.4 μmol/l, ng DNA, 1 U Taq DNA ( ) PE 9700 PCR PCR PCR 6% (Acr.. Bis=19.. 1, )PCR 1h, 95 PCR 10 min, Ladder [4], 40 W, 2 3 h 10%, 1%, 3%, 10%, Ladder 1.5, SPSS13.0 (Heterozygosity, H), (Discrimination

3 9 : X-STR 1145 表 1 9 个 X-STR 基因座引物序列及扩增条件 Table 1 Primer sequences and PCR conditions of 9 X-STR loci Loci DXS7130 DXS7132 DXS6804 DXS8378 DXS7424 DXS7423 DXS6789 DXS6799 HPRTB Primer s sequence(5 3 ) Forward: CTGCAAGCCATTTGGAATAT Reverse: TCCTAGGACTGGGAAAGGAC Forward: AGCCCATTTTCATAATAAATCC Reverse: AATCAGTGCTTTCTGTACTATTGG Forward: CCCAGATATTTTGACCACCA Reverse: GGCATGTGGTTGCTATAACC Forward: CACAGGAGGTTTGACCTGTT Reverse: AACTGAGATGGTGCCACTGA Forward: CTGCTTGAGTCCAGGAATTCAA Reverse: GAACACGCACATTTGAGAACATA Forward: TAGCTTAGCGCCTGGCACATA Reverse: GTCTTCCTGTCATCTCCCAAC Forward: TTGGTACTTAATAAACCCTCTTTT Reverse: CTAGAGGGACAGAACCAATAGG Forward: ATGAATTCAGAATTATCCTCATACC Reverse: GAACCAACCTGCTTTTCTGA Forward: ATGCCACAGATAATACACATCCCC Reverse: CTCTCCAGAATAGTTAGATGTAGG PCR conditions s s s s s s 32cycles s s s s s s s s s s s s 32 cycles s s s s s s s s s power of female, PD F ), (Discrimination power of male, PD M ), (Polymorphism information content, PIC), (Probability exclusion, PE) (Haplotype diversity, HD)=(n/n 1)(1 Pi 2 )[Pii, n ] χ 2 Hardy-Weinberg Phylip3.65 Nei [5], MEGA 4.0 (UPGMA ), SPSS , DXS7130 DXS7132 DXS6804 DXS7423 DXS7424 DXS6789 DXS6799 DXS8378 HPRTB 9 X-STR , , DXS7130 DXS7132 DXS6804 DXS7423 DXS7424 DXS6789 DXS6799 DXS8378 HPRTB 9 X-STR , 3χ 2, Hardy-Weinberg (P>0.05) X-STR 4, (H) (PIC)(PD F ) (PD M ) (PE) (PE) 2.4 DXS7130 DXS , DXS6789 DXS6799 DXS7424 DXS , ( 5 6) X-STR [6~11], χ 2 7(P 0.05),,, DXS6789,,

4 1146 HEREDITAS (Beijing) 表 2 甘肃裕固族 9 个 X-STR 基因座等位基因频率 Table 2 Allele frequencies of nine X-STR in Gansu Yugur ethnic group DXS7130 DXS7132 DXS6804 DXS8378 DXS6799 Allele Female Male Female Male Female Male Female Male Female DXS7423 DXS7424 DXS6789 HPRTB Allele Female Male Female Male Female Male Female Male Male X-STR (DXS7423, DXS8378, DXS7132, DXS6789, DXS6799 DXS6804), (Yugur) (Xi an Han) (Hui) (Tibetan) (Uygur) (Mongolian) (Ewenki) (Oroqen) (Daur) [6~14] 9 Nei, 8, Nei 1 2

5 9 : X-STR 1147 表 3 9 个 X-STR 基因座在裕固族女性中基因型的分布 Table 3 s of nine X-STR loci in female Yugur group DXS7130 DXS7132 DXS6804 DXS7424 DXS Total P= P= P= P= P= DXS7423 DXS8378 DXS6799 HPRTB Total P= P= P= P= 表 4 甘肃裕固族群体 9 个 X-STR 基因座群体遗传多态性指标 Table 4 Polymorphic parameters of nine X-STR loci in Gansu Yugur ethnic group Loci H PIC PD F PD M PE (trios) PE (diad) DXS DXS DXS DXS DXS DXS DXS DXS HPRTB Combined >

6 1148 HEREDITAS (Beijing) 表 5 裕固族男性 DXS7130 和 DXS8378 两个基因座单体型频率 (n=65) Table 5 Haplotype frequencies of the loci DXS7130 and DXS8378 in males of Yugur ethnic group (n=65) Haplotype Frequency Haplotype Frequency 1 11/ / / / / / / / / / / / / / / HD: = HD: Haplotype diversity= 表 6 裕固族男性 DXS6789 DXS6799 DXS7424 和 DXS6804 等 4 个基因座单体型频率 (n=65) Table 6 Haplotype frequencies of the loci DXS6789, DXS6799, DXS7424, and DXS6804 in males of Yugur ethnic group (n=65) Haplotype Frequency Haplotype Frequency 1 18/11/15/ /11/15/ /11/17/ /11/16/ /12/14/ /11/16/ /10/17/ /11/16/ /10/17/ /11/17/ /11/15/ /11/17/ /11/16/ /11/19/ /11/16/ /12/16/ /11/16/ /12/16/ /11/17/ /12/18/ /11/17/ /9/14/ /11/18/ /9/17/ /11/18/ /10/16/ /11/18/ /11/15/ /12/14/ /11/16/ /12/16/ /11/16/ /13/17/ /11/17/ /11/17/ /11/17/ /11/18/ /11/18/ /10/18/ /12/15/ /11/15/ /12/16/ /11/16/ /9/17/ /11/17/ /9/17/ /12/16/ /11/17/ /13/17/ /11/17/ /10/17/ /11/18/ /10/17/ /11/18/ /11/14/ HD: = HD: Haplotype diversity=

7 9 : X-STR 1149 表 7 裕固族与其他人群 X-STR 基因座遗传差异比较 Table 7 The comparison of X-STR heredity with Yugur and other groups Loci Xi an Han Mongol Tibetan Lhoba Ewenki DXS DXS DXS DXS DXS DXS DXS DXS HPRTB X-STR, P<0.05 means there is no X-STR population data at present, P<0.05. 表 8 9 个群体的遗传距离 Table 8 The genetic distance of 9 populations based on 6 X-STR loci Achang Population Yugur Xi'an han Hui Tibetan Uygur Mongol Ewenki Oroqen Xi an han Hui Tibetan Uygur Mongol Ewenki Oroqen Daur 图 1 9 个群体的系统进化树 Fig. 1 Phylogeny tree of 9 populations 图 2 9 个群体的聚类分析结果 Fig. 2 Clustering results of 9 populations

8 1150 HEREDITAS (Beijing) (Short tandem repeat, STR),,,, Y STR, X-STR [14,15], X [16],,, [2] 9 X-STR, X, 4, DXS7424,, PCR,,,,, 9 X-STR 54, ~0.6909, DXS8378 4, DXS7423 5, DXS7132 DXS6804 DXS6799 DXS7424 HPRTB 6, DXS6789 7, DXS χ 2, 9 X-STR Hardy-Weinberg (P>0.05) (H)(PIC)(DP) (PE),,, PIC>0.5,, 0.5>PIC>0.25,, PIC<0.25,, DP>0.8, PE>0.5,,, X-STR, DXS7423 DXS8378 ( ), ( ), ( , ), H ~0.8096, PIC , DP , DXS X-STR, DXS7423 DXS8378,, 9 X-STR PIC , PD F , PD M , PE , PE ,, X-STR, 9 X-STR PD F PD M, X-STR,, 10 Mb, [17], XSTR, [18] Xp22 q28 STR, 4 9 X-STR 4, DXS7130 DXS8378 ; DXS7132 DXS7424 DXS6789 DXS6799 DXS6804 ; HPRTB ; DXS7423 DXS7130 DXS8378, X ( Xp ) 4.15 Mb 8.78 Mb [18] DXS6789( cm) DXS6799( cm) DXS7424(104.9~121 cm) DXS6804( cm) [15],, [19],, DXS7130 DXS8378 DXS6789 DXS6799 DXS7424 DXS6804, DXS7130 DXS , DXS6789 DXS6799 DXS7424 DXS , ,,

9 9 : X-STR 1151 [20],,, DXS6789,,, STR,,, 2000, ,,, [21],, [22],,,, 6 X-STR 9,,, ,, , ;, Nei, 9,,,,, [23],,, Shriver [24], STR,, X STR 9 6 X STR,, X-STR DXS7423 DXS X-STR,,,,,, DNAX Y,,,, (References): [1] The International HapMap Consortium. A haplotype map of the human genome. Nature, 2005, 437(7063): [DOI] [2] CHEN Yan-Jiong, CHEN Teng, LAI Jiang-Hua, ZHENG Hai-Bo, WANG Zhan-Hai, JIN Tian-Bo, LI Sheng-Bin. Genetic polymorphisms of 9 STR loci in Yugu ethnic group in Gansu Province. Journal of Xi an Jiaotong University (Medical Sciences), 2003, 24(3): ,,,,,, 9 STR. ( ), 2003, 24(3): [3] Walsh BS, Petzger DA, Higuchi R. Chelex-100 as medium for simple extraction of DNA for PCR-based typing from forensic material. Biotechnics, 1991, 10(6): [4] YU Bing, QIN Qun-Xia, YAN Jin-Cheng, ZHANG Hong-Bo, LI Sheng-Bin. Genetic polymorphism analysis of 6 STR loci on the X chromosome in Xi an Han population. Journal of Forensic Medicine, 2005, 21(3): ,,,,. X 6 STR., 2005, 21(3): [5] Takezaki N, Nei M. Genetic distances and reconstruction of phylogenetic trees from microsatellite DNA. Genetics, 1996, 144(1): [6] YU Bing, ZHANG Hong-Bo, LI Sheng-Bin. X chromosome STRs polymorphisms of Han ethnic group from Northwest China. Forensic Science International, 2005, 153: [DOI] [7] LIU Qing-Bo, LI Sheng-Bin. Patterns of genetic polymorphism at the 10 X-chromosome STR loci in Mongol population. Forensic Science International, 2006, 158:

10 1152 HEREDITAS (Beijing) [DOI] [8] GAO Ya, JIN Tian-Bo, YU Bing, DU Hong, LI Sheng-Bin. Genetic polymorphisms of 10 X chromosome STR loci in a Chinese Tibetan population. Chin J Med Genet, 2006, 23(1): ,,,, X 10 STR, 2006, 23(1): [9] KANG Long-Li, LI Sheng-Bin. X-chromosome STR polymorphism of Luoba ethnic group living in Tibet (SW China). Forensic Science International, 2006, 156: [DOI] [10] GU Shan-Zhi, LI Sheng-Bin. X-chromosome STRs analysis of Ewenke ethnic population. Forensic Science International, 2006, 158: [DOI] [11] GU Shan-Zhi, ZHONG Jing-Fang, MU Zhi-Qiang, LI Sheng-Bin. Polymorphism of five X-STR loci in Chinese Achang ethnic group. J Zhejiang Univ (Medical Sci), 2007, 36 (2): ,,,. 5 X-STR ( ), 2007, 36(2): [12] FENG Xue, XU Ping, TUO Ya, YU Bing, LI Sheng-Bin. An investigation for genetic polymorphisms of 10 STR loci on chromosome X of Chinese Hui nationality population in Ningxia region of China. Chinese Journal of Medical Genetics, 2006, 23(3): ,,,,. X 10., 2006, 23(3): [13] HOU Qiao-Fang, YU Bin, LI Sheng-Bin. Genetic polymorphisms of nine X-STR loci in four population groups from Inner Mongolia, China. Genomics, Proteomics & Bioinformatics, 2007, 5(1): [DOI] [14] REN Yun-Xing, LIANG Jing-Qing. Research progress on X chromosome STR loci in forensic medicine. Hereditas (Beijing), 2006, 28(10): ,. X STR., 2006, 28(10): [15] Edelmann J, Hering S, Kuhlisch E, Szibor R. Validation of the STR DXS7424 and the linkage situation on the X-chromosome. Forensic Sci Int, 2002, 125(2-3): [DOI] [16] WU Ling-Qian, PAN Qian, LONG Zhi-Gao, ZHU Jun-Zhen, DAI He-Ping, ZHENG Duo, XIA Kuen, HUANG Xing-Qing, XIA Jia-Hui. Genetic diagnosis and prenatal genetic diagnosis of fragile X syndrome. Hereditas (Beijing), 2003, 25(2): ,,,,,,,,. X, 2003, 25(2): [17] Szibor R, Krawczak M, Hering S, Edelmann J, Kuhlisch E, Krause D. Use of X-linked markers for forensic purposes. Int J Legal Med, 2003, 117(2): [18] LV De-Jian. Detecting haplotypes of DXS7132 and DXS6804 loci by multiplex PCR. Acta Genetica Sinica, 2003, 30(1): PCR DXS7132 DXS6804., 2003, 30(1): [19] Szibor R, Krawczak M, Hering S, Edelmann J, Kuhlisch E, Krause D. Use of X-linked markers for forensic purposes. Int J Legal Med, 2003, 117: [20] Reinhard Szibor, Sandra Hering, Eberhard Kuhlisch, Ines Plate, Susanne Demberger, Michael Krawczak, Jeanett Edelmann. Haplotyping of STR cluster DXS6801 DXS6809 DXS6789 on Xq21 provides a powerful tool for kinship testing. Int J Legal Med, 2005, 119: [DOI] [21] DU Ruo-Fu. Chinese Population Genetics. Beijing: Science Press, :, [22] GAO Zi-Hou. The discourse of two subdivisions of Yugu ethnic group origin and development. North West Ethno-national Studies, 1995, (16)1: , 1995, (16)1: [23] LI Tian-Xue. On the phase of sub-ethnic consciousness about formation of the Yugurs. Nationalities Research in Qinghai, 2005, 16(2): , 2005, 16(2): [24] Shriver MD, Jin L, Boerwinkle E, Deka R, Ferrell RE, Chakraborty R. A novel measure of genetic distance for highly polymorphic tandem repeat loci. Molecular Biology And Evolution, 1995, 12(5):

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Microsoft Word _new_.doc HEREDITAS (Beijing) 2009 2, 31(2): 153 159 ISSN 0253-9772 www.chinagene.cn 研究报告 DOI: 10.3724/SP.J.1005.2009.00153 云南白族傣族彝族 X 染色体 STR 基因座的遗传多态性及其与中国 5 个主要民族的遗传关系魏曙光, 杨丽, 郑海波, 沈靓, 赖江华,, 710061 应用复合 PCR