大熊猫遗传多样性评估的微卫星分型体系优化

Optimization of microsatellite genotyping system used for genetic diversity evaluation of Ailuropoda melanoleuca

微卫星作为重要的分子标记之一,已被证明在大熊猫种群规模评估、亲子鉴定和遗传多样性分析方面是有效的。目前微卫星标记在大熊猫(Ailuropoda melanoleuca)染色体上物理定位方面的报道较少,而且缺乏微卫星基因分型系统的效能评估以及PCR扩增条件的优化。本研究基于大熊猫基因组参考序列(ASM200744v2),分析了34个大熊猫微卫星位点的染色体定位特征并评价了位点的应用价值。通过优化34个STR-PCR反应体系和扩增程序,结合微卫星的染色体定位数据确定了Ame-μ10标记的较低应用价值以及gpz-6重新筛选引物的必要性。本研究有助于提高基因分型结果的重复性和可靠性,对促进《大熊猫种群遗传档案建立技术规程》规范化应用和制定大熊猫保护策略具有重要意义。

Microsatellite DNA is one of the most widely used genetic markers of giant panda,especially in population size estimation,paternity testing,and genetic diversity analysis.However,there are few reports on the physical locations of microsatellite markers on the chromosomes of the giant panda(Ailuropoda melanoleuca)and research on the performance of microsatellite in genotyping system and the PCR amplification conditions.In this study,we analyzed the chromosomal locations and evaluated the application values of 34 microsatellite markers,based on the giant panda genome reference sequence(ASM200744v2).We optimized the PCR reaction systems and amplification procedures for these 34 microsatellite markers.We found the low value of the microsatellite marker of Ame-μ10 in genetic application,and the necessity in redesigning the primers for gpz-6.Our research helps to improve the reproducibility and reliability of genotyping results and is of great significance for promoting the establishment and standardized application of the“A Regulation for Giant Panda Population Genetic Archives”in giant panda conservation.