卤虫(AC)_n和(AG)_n微卫星文库构建

Construction of (AC)_n and (AG)_n Microsatellite DNA Libraries in Brine Shrimp Artemia

刘海珍¹, 邹小艳², 郭松长², 李双¹, 张湑泽³, 马建滨^1*, 都玉蓉^1*

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作者单位：1. 青海师范大学生命与地理科学学院, 青藏高原环境与资源教育部重点实验室, 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 810007;
3. 青海民族大学, 西宁 810001

中文关键字：卤虫;微卫星;富集文库

英文关键字：Artemia; microsatellite; enriched library

中文摘要：卤虫Artemia作为重要的基础实验材料和经济动物资源,其微卫星位点具有重要的研究和应用价值。本研究使用磁珠富集法,成功构建了卤虫的(AC)_n和(AG)_n微卫星富集文库。通过对文库进行PCR方法鉴定和测序确认,所构建的(AC)_n和(AG)_n文库阳性克隆率分别为55.8%和34.5%。依据得到的微卫星序列,设计并合成了63对微卫星引物,随机以2个地理单元(青海、西藏)共18个卤虫基因组DNA为扩增模板,筛选出6对具有多态性的卤虫微卫星引物。本研究筛选出的微卫星多态位点可用于卤虫种群的遗传多样性评估、遗传图谱的构建和数量性状定位等后续工作。

英文摘要：Brine shrimp Artemia is an important model organism and economic animal resource. The microsatellite loci of this species have a significant research and application value. In this study, the Artemia microsatellite enriched DNA libraries of (AC)_n and (AG)_n were conducted successfully using the method of FIASCO (fast isolation by AFLP sequence containing repeats). In the (AC)_n library, 67 colonies (total no. 120) were picked out and sequenced, with a positive clone rate of 55.8%. In the (AG)_n library, 69 colonies out of 200 colonies were sequenced, with a positive clone rate of 34.5%. Among all the sequences containing microsatellites, 63 pairs of primers were designed and synthesized for further primer screening. By using 18Artemia DNA samples from Qinghai and Tibetan, 6 pairs of primers with polymorphic microsatellite loci of Artemia were identified. These polymorphic loci may facilitate further study on the detection of genetic diversity, genetic linkage mapping and the location of quantitative traits of Artemia.

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