大熊猫微卫星DNA的筛选及其应用
Microsatellite Dnas and Kinship Identification of Giant Panda
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摘要: 经DpnⅡ限制酶消化大熊猫基因组DNA后,选取150—500 bp大小的片段构建了DNA文库。用合成的(CA)15探针从这一文库中克隆筛选了10个大熊猫特异的微卫星DNA座位。在测定DNA序列的基础上设计并合成了10个座位特异的引物,以PCR技术扩增了7份抽提自组织细胞和6份抽提自毛发的大熊猫DNA样品。发现除座位g007外,其余9个座位均呈多态性,而且所有座位均为偶数碱基的长度变异。对有准确谱系记录的家系的研究结果证明,这10个微卫星DNA座位严格按孟德尔方式遗传。因而这些座位能有效地应用于大熊猫的亲子鉴定,从而为建立各地大熊猫谱系和制定有效的繁殖计划提供了一个有效的遗传学手段。根据这10个微卫星座位的分析,我们澄清了两组未知的父系关系。Abstract: We constructed a DNA library of 150-500 bp fragments from giant panda genomic DNA digested with DpnⅡ.Ten microsatellite DNA loci were isolated and characterized by screening the library with a 32P-labelled (CA)15 probe.The specific primers for each of the microsatellite DNA locus were synthesized and employed to amplify DNA samples isolated from 7 specimens of panda tissue and 6 specimens of hair.Nine of the 10 microsatellite DNA loci were polymorphic in the 13 samples.Inheritance patterns of alleles at these loci accurately agree with the well recorded pedigree.The polymorphism on these loci also clarified some previously unresolved paternity.These results imply the screened microsatellite DNA loci are useful makers for the identification of kinship among giant panda.