藏羚羊(AC)n, (AG)n微卫星文库构建
Construction of (AC)n and (AG)n Microsatellite DNA Libraries in Pantholops hodgsonii
都玉蓉1，马建滨1，徐永涛1，周智红1，谭春敏1，张湑泽2，郭松长2,3*
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DOI：
作者单位：(1. 青海师范大学生命与地理科学学院, 西宁 810008；2. 中国科学院高原生物适应与进化重点实验室, 西宁 810001；3. 青海省作物分子育种重点实验室, 西宁 810007)
中文关键字：藏羚羊；微卫星；富集文库
英文关键字：Pantholops hodgsonii; microsatellite; enriched-library
中文摘要：藏羚羊Pantholops hodgsonii是我国珍稀的动物资源，开发其微卫星标记具有重要的科研与应用价值。本研究采用FIASCO法(Fast Isolation by AFLP Sequences Containing repeats)构建了藏羚羊的(AC)n和(AG)n微卫星富集文库。各文库随机挑取150个白色菌落，分别筛选到76个(AC)n和61个(AG)n阳性克隆；经测序得到含AC重复的微卫星序列37条和含AG重复的序列29条。研究共设计、合成微卫星引物47对，以扩增、电泳等方法进行筛选，最终获得15个可稳定扩增且具有多态的微卫星标记。筛选出的引物可用于评估藏羚羊的核DNA遗传多样性、遗传结构、种群动态以及构建藏羚羊遗传图谱等。
英文摘要：Tibetan antelope (chiru, Pantholops hodgsonii) is an indigenous species of Qinghai-Tibetan Plateau and listed as an AppendixⅠspecies in CITES and Category Ⅰ in the Key Protected Wildlife List of China. Microsatellites and SSR are powerful molecular markers of numerous biological contexts. In this study, the microsatellite enriched-libraries of the chirus were constructed according to the FIASCO (Fast Isolation by AFLP of Sequences Containing repeats) protocol. Finally, 76 and 61 positive clones were screened in 150 white clones picked from (AC)n and (AG)n microsatellite libraries, respectively. Based on the result of sequencing, 37 SSR were detected from (AC)n library and 29 from (AG)n library among the positive clones. A total of 15 microsatellite loci were identified to be polymorphic using 47 designed primer pairs. These may facilitate further studies on the genetic diversity, genetic structure, demographic history and genetic linkage mapping of Tibetan antelopes.