摘要
通过磁珠富集法构建了藏酋猴AC重复和AAAG重复的微卫星富集文库,分离微卫星序列并对其进行分析。将藏酋猴基因组DNA经Sau3AI酶切后纯化回收,连接特定接头。用生物素标记的探针与酶切片段杂交,捕获300～1000bp片段,随后将获得的片段连接到pMD-19T载体上,转化至JM109中,成功构建藏酋猴微卫星富集文库。(AC)n富集文库和(AAAG)n富集文库的阳性克隆率分别为50%和10%左右。根据测序得到的48个微卫星序列成功设计了24对引物,最终筛选出6个微卫星标记,这些标记将为藏酋猴的遗传多样性研究、圈养种群结构的分析和遗传图谱的构建等奠定一定的基础。
Microsatellite-enriched libraries of AC-repeats and AAAG-repeats in Macaca thibetana were constructed using an enrichment method with biotin-labeled probes and streptavidin-coated magnetic beads.The ratios of positive clones containing（AC）n and（AAAG）n enriched libraries were approximately 50% and 10%,respectively.Total 48 microsatellite sequences were obtained by sequencing positive clones.According to sequencing results,24 pairs of primers were designed,and of them 6 markers were optimized.The 6 microsatellite makers of M.thibetana obtained from this study would be significant in future study,such as population structure analyses,paternity testing,and genetic mapping.
出处
《四川动物》
CSCD
北大核心
2012年第1期39-44,共6页
Sichuan Journal of Zoology
基金
成都大熊猫繁殖研究基金
关键词
藏酋猴
富集文库
微卫星
Macaca thibetana
enriched library
microsatellite
