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利用微卫星DNA分型对韩国纯血马进行亲子鉴定

Parentage testing of Thoroughbred horse in Korea using microsatellite DNA typing.

作者信息

Lee Sun-young, Cho Gil-jae

机构信息

Laboratory of Equine Genetics, Korea Racing Association, Gwacheon 427-711, Korea.

出版信息

J Vet Sci. 2006 Mar;7(1):63-7. doi: 10.4142/jvs.2006.7.1.63.

Abstract

The present study was to construct a parentage testing system for Thoroughbred (TB) horse. A total number of 1,285 TB horse samples including 962 foals for parentage testing, 9 sires and 314 dams for individual identification were genotyped. Genomic DNA was extracted from 5 hair roots and genotyped by using 14 microsatellite markers (AHT4, AHT5, ASB2, ASB17, ASB23, CA425, HMS1, HMS3, HMS6, HMS7, HTG4, HTG10, LEX3 and VHL20). This method consisted of multiplexing PCR procedure and showed reasonable amplification of all PCR products. Genotypes were determined by genetic analyzer. The number of alleles per locus varied from 3 to 9 with a mean value of 6.36 in TB horse. The expected heterozygosity was ranged from 0.548 to 0.831 (mean 0.699), and the total exclusion probability of 14 microstellite loci was 0.9998. Of the 14 markers, ASB2, ASB17, ASB23, HMS7 and HTG10 loci have relatively high PIC value (> 0.7). Of the 962 foals, 960 foals were qualified by compatibility according to the Mendelism. These results suggest that the DNA typing method has high potential for parentage verification and individual identification of TB horses.

摘要

本研究旨在构建纯血马亲子鉴定系统。对总共1285份纯血马样本进行基因分型,其中包括962份用于亲子鉴定的幼驹样本、9份种公马样本和314份母马样本用于个体识别。从5根毛发根部提取基因组DNA,并使用14个微卫星标记(AHT4、AHT5、ASB2、ASB17、ASB23、CA425、HMS1、HMS3、HMS6、HMS7、HTG4、HTG10、LEX3和VHL20)进行基因分型。该方法包括多重PCR程序,所有PCR产物均显示出合理的扩增效果。通过基因分析仪确定基因型。纯血马中每个位点的等位基因数量从3到9不等,平均值为6.36。预期杂合度范围为0.548至0.831(平均0.699),14个微卫星位点的总排除概率为0.9998。在14个标记中,ASB2、ASB17、ASB23、HMS7和HTG10位点具有相对较高的多态信息含量值(>0.7)。在962匹幼驹中,有960匹幼驹根据孟德尔遗传定律符合亲子关系匹配。这些结果表明,该DNA分型方法在纯血马的亲子关系验证和个体识别方面具有很高的潜力。

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