果子狸多态性微卫星位点的筛选及特性分析

以采自四川平武县老河沟自然保护区的一份果子狸(Paguma larvata)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...

Development and Characterization of Polymorphic Microsatellite Markers in the Masked Palm Civet (Paguma Larvata)

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 10⁹unrelated individuals or 10⁴ 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.