286 HEREDITAS (Beijing) ,, ;,, (Short tandem repeats, STR), DNA,, [1] 1996 DNA [2] 9 3 PCR [3] [4, 5], [6] ABI Bovine Paternity PCR Typing Kit(

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1 HEREDITAS (Beijing) , 31(3): ISSN 研究报告 DOI: /SP.J 微卫星在种公牛个体识别与亲缘鉴定方面的应用王静 1, 刘丑生 2, 张利平 3, 王志刚 2, 于福清 2, 张桂香 2, 孙秀柱 2, 韩旭 2 4, 魏玉春 1., ; 2., ; 3., ; 4., 采用美国 ABI 公司牛亲子鉴定试剂盒 (Bovine Paternity PCR Typing Kit, 包括 11 个常染色体 ) 和 3 个自选的 Y 染色体微卫星座位, 检测我国部分种公牛站肉用种公牛 14 个微卫星座位的多态性分布, 评估其遗传多样性, 并探讨其用于个体识别与亲缘鉴定的可行性结果表明 : 种公牛在 14 个微卫星座位中遗传多样性均较高, 其中 MCM158 座位的平均多态信息含量最高达到 0.888, ETH10 座位的最低, 为单个座位的个体识别能力在 0.715~0.968 之间, 累积个体识别能力为 99.99%, 累计非父排除率达到 99.99%, 表明采用的 14 个位点适用于个体识别和亲缘鉴定种公牛 ; 微卫星 ; 个体识别 ; 亲缘鉴定 Individual identification and paternity testing of bulls using microsatellite WANG Jing 1, LIU Chou-Sheng 2, ZHANG Li-Ping 3, WANG Zhi-Gang 2, YU Fu-Qing 2, ZHANG Gui-Xiang 2, SONG Xiu-Zhu 2, HAN Xu 2, WEI Yu-Chun 4 1. Lanzhou Vocational Technology College, Lanzhou , China; 2. National Center of Preservation and Utilization of Genetic Resources of Domestic Animals and Forage, National Animal Husbandry and Veterinary Service, Beijing , China; 3. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou , China; 4. College of Animal Science and Technology, Northwest Sci-Tech University of Agriculture & Forestry, Yangling , China Abstract: The polymorphism distributions of 14 microsatellite loci were detected using the Bovine Paternity PCR Typing Kit (including11 X-STR) and 3 selected Y-STR microsatellite DNA markers. The genetic diversity were evaluated, and the feasibility of the application to individual identification and paternity testing were discussed. The results showed that all the 14 microsatellite loci had genetic polymorphisms in bulls, and the polymorphism information content (PIC) in loci MCM158 was the biggest (0.888), while the ETH10 was the lowest (0.482). Power of discrimination (DP) value of the 14 STR loci ranged from to The Cumulate DP (CDP) was 99.99%, and the Cumulate PE (CPE) also reached 99.99%. These results indicate that the 14 microsatellites can be applied to the individual identification. Keywords: bull; microsatellite;individual identification; paternity testing 收稿日期 : ; 修回日期 : 基金项目 : ( [2007]43 ) 作者简介 : (1982 ),,, Tel: ; jwang_423@126.com 通讯作者 : (1964 ),,, liuchousheng@sina.com

2 286 HEREDITAS (Beijing) ,, ;,, (Short tandem repeats, STR), DNA,, [1] 1996 DNA [2] 9 3 PCR [3] [4, 5], [6] ABI Bovine Paternity PCR Typing Kit( 11 ), 3 Y, 14,,, 1 材料和方法 (, 80, 20 ) 10 ml/, 基因组 DNA 准备 DNA 1.5%, 100 ng/μl PCR 引物选择 11 (Bovine Paternity PCR Typing Kit, ABI ) [7, 8] MCM158 MAF45 UMN Y, 14 Y 5 TET 6FAM, PCR 扩增,, (Bovine Paternity PCR Typing Kit, ABI ) Y PCR 25 μl, DNA 100 ng, 0.15~0.30 μmol/l, 1 U Taq, 0.2 mmol/l dntps, 1~1.5 mmol/l MgCl 2 1 PCR buffer PCR : 95 4 min; s, 56~65 45 s, 72 1 min, 30~35, min, PCR 产物检测 PCR 8%,, PCR ABI3100-Avant, ABI3100-Avant, LIZ500, Gene-MapperVersion 统计分析, ABI3100-Avant, GENEPOP Version3.4 (A) (H o ) (H e ) [9] [10] (PIC) PowerStats V1.2(Promega Corporation,U.S.A.) (DP) (PM) [11], Cervus2.0 (PE) [12] 2 结果与分析 2.1 DNA 100,

3 3 : ( 1) Y 3 ( 2), , 9.8 TGLA122, 19, BM1824, , ( 1) 1 14 MCM , ETH10, ,, 1 (H o ) (H e ) 0.84~ ~ , ~0.968, MCM , ETH10, MCM158, ETH10,,, %, (PM) 图 1 常染色体上 11 个座位毛细管电泳检测图 (Marker) X, Y 图 2 Y 染色体上个座位毛细管电泳检测图 (Marker) X, Y

4 288 HEREDITAS (Beijing) 表 1 14 个微卫星座位上的信息 (PIC) (He) (Ho) (DP) (PE) BM BM ETH ETH ETH INRA MAF MCM TGLA TGLA TGLA TGLA UMN SPS , ~0.694, TGLA , ETH10, %, 3 讨论 3.1 (PIC) Botstein [10] : PIC>0.5, ; 0.25<PIC<0.5, ; PIC<0.25, 14 (ETH10), : 0.72, Takezaki [13] 0.3~0.8, ~0.968, 99.99%, (PM) 10 9, [14],, [6] %,,,, 20 60,,,,,,,,,, ~0.694,

5 3 : %, ( ),, ( ), 14, Ellegren [15], 5 ( 6 ) 98%, % [16] 6 STR STR, , , 99.99% ABI 11, 3 Y,,, ,, DNA,,, SPSS13.0,,, 参考文献 (References): [1].. :, 2001, [2], Galli Andrea,.., 2002, 3: [3]. [ ]. :, [4],,,,. 6 STR., 2004, 26(3): [5] Schnabel RD, Ward TJ, Derr JN. Validation of 15 microsatellites for parentage testing in North American bison, Bison bison and domestic cattle. Anim Genet, 2000, 31(6): [6],,,,,,. STR., 2005, 21(2): 4 6. [7] Liu WS, Mariani P, Beattie CW. A radiation hybrid map for the bovine Y Chromosome. Mammalian Genome, 2002, 13(6): [8],. SRY., 2006, 28(2): [9] Nei M. Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics, 1978, 89(3): [10] Botstein D, White RL, Skolnick M, Davis RW. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. American Journal of Human Genetics, 1980, 32(3): [11] Tereba A. Tools for analysis of population statistics, Profiles DNA, 1999, (2): [12] Garber RA, Morris JW. General Equations for the Average Power of Exclusion for Genetic Systems of Nco-dominant Alleles in One-parent Cases of Disputed Parentage. In: Walker RH,Ed. Inclusion Probabilities in Parentage Testing, American Association of Blood Banks. Arlington, VA, USA, 1983, [13] Takezaki N, Nei M. Genetic distances and reconstruction of phylogenetic trees from microsatellite DNA. Genetics, 1996, 144(1): [14],,,,,. DNA., 2001, (5): 4 7. [15] Ellegren H, Johansson M, Sandberk K.Cloning of highlypoly morphic micosatellite in the horse. Anim Genetics, 1992, (23): [16],,,,,,,,,,. DNA., 2005, 13(9):

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