摘要
采用磁珠富集法构建了红砂（Reaumuria soongarica）基因组微卫星富集文库,通过测序结果对文库的特性进行了分析。结果表明：在获得的13 922条微卫星序列中完美型占88%,非完美型10%,混合型2%。微卫星重复基元显示5种类型,以三碱基基元（GGT）n、（TGG）n、（ACC）n最多,占总数的60.7%;其次为二碱基基元,主要是（AC）n、（GT）n、（CA）n,占38.6%;这5种微卫星重复基元的重复次数多集中在5～10。通过对随机挑选的100条微卫星序列设计引物,并以遗传距离和地理距离较远个体的基因组为模板进行扩增筛选,可有效扩增的引物为62对,并进一步测序筛选获得了12对多态性微卫星引物。磁珠富集法是开发红砂基因组微卫星序列的一种高效、可行的方法,构建的微卫星文库效率较高,富集量大,重复基元类型丰富,为红砂遗传进化和种质资源保护提供了可靠的数据支持和高效的分子遗传标记。
The present study used the method of magnetic beads enrichment to build enriched microsatellite sequence library and analyzed the characteristics of library through sequencing results.The results showed that13 922 microsatellite sequences were obtained with 88% perfect,8%imperfect and 2% compound repeat motifs.Of these microsatellite loci,five type repeat primitives were presented,including prominent 60.7%three base primitive with（GGT）n,（TGG）n and（ACC）n,and 38.6%two base primitive with（AC）n,（GT）n and（CA）n.And the repeat time was mainly between 5 and 10,and declined as exceeding 30 times.Total 62 pairs of efficient PCR primers and 12 pairs of microsatellite primers have been screened by designing the primers with the 100 random microsatellite sequence and PCR screening with templates in populations with distant genetic and physical distance.Here,we revealed that the method of magnetic beads enrichment was high efficient and feasible for the development of Reaumuria soongarica genome microsatellite sequence.The present investigation would provide informative support and efficient genetic molecular marker for the genetic evolution and germplasm resources protection of Reaumuriasoongarica.
出处
《中国沙漠》
CSCD
北大核心
2016年第2期374-382,共9页
Journal of Desert Research
基金
中国科学院"百人计划"项目(29Y127E71)
国家自然科学基金项目(31370395
31500266)
