【摘要】 以采自四川平武县老河沟自然保护区的一份果子狸(Pagumalarvata)组织样品为实验材料,通过FIASCO法构建微卫星文库,对250个单克隆进行测序分析,获得147条含微卫星序列的片段,微卫星单元重复次数大于10的序列共有42条。据此设计引物42对,进一步使用21份果子狸样品检测这些引物的扩增能力和扩增序列多态性,最终得到5个多态性微卫星位点。同时,对已发表的13个果子狸微卫星位点进行检测,发现其中5个位点具有多态性。对这10个位点的特性进行分析,结果显示它们具有较高的多态性(等位基因数为2～11,观测杂合度为0.286～0.737,期望杂合度为0.358～0.906);PID和PID-sib值表明,在约10~9个没有亲缘关系的个体或10~4个有亲缘关系的个体中,可能发现一对基因型相同的个体,由于果子狸野外局域种群规模远低于这个量级,这10个微卫星位点可用于果子狸的个体识别。更多还原

【Abstract】 To provide genetic tools for individual identification of the masked palm civet(Paguma larvata) through DNA analysis, we selected a tissue sample from Laohegou Nature Reserve in Sichuan for constructing microsatellite library by FIASCO. Altogether 250 clones were sequenced and 147 sequences were found containing microsatellite motifs, in which there were 42 sequences with >10 motif repeats. Primer pairs were designed based on these 42 sequences. Through tests for amplification and polymorphism using additional 21 samples of masked palm civets, 5 polymorphic microsatellite markers were obtained. In the meantime, 13 microsatellite markers published previously were tested for the masked palm civet, and found 5 of them polymorphic. Collectively, these 10 markers showed relatively high polymorphism(number of alleles was 2–11, observed heterozygosity was 0.286–0.737, and expected heterozygosity was 0.358–0.906). The values of PID and PID-sib indicated that there would be individuals with completely identical genotypes in about 10~9 unrelated individuals or 10~4 related individuals. As the size of wild populations in a particular region was greatly lower than this number, these 10 microsatellite markers were sufficient for individual identification for the masked palm civet.更多还原