基于绵羊皮肤组织ESTs开发新型微卫星标记.pdf

中国农学通报 2010,26(3):42-45 Chinese Agricultural Science Bulletin 基于绵羊皮肤组织ESTs开发新型微卫星标记 王遵宝，努尔力·阿不力孜，赵宗胜，曾献存，吴洪宾 （石河子大学动物科技学院，新疆石河子832003） 摘 要：利用生物信息学方法，在从国际公共生物数据库检索获得绵羊皮肤组织相关的3295 条EST 序列 中筛选微卫星标记。结果表明，从绵羊皮肤组织相关的EST 序列中搜索到微卫星209 个，含有微卫星的 序列196 个，占整个EST 序列数据库的6.3% ，其中双碱基重复150 个，三碱基重复19 个，四碱基重复12 个，五碱基重复17 个，六碱基重复11 个。在这些微卫星序列中，AC/TG 重复在双碱基类型中最丰富，占 双碱基微卫星序列总数的67%。根据筛选到的微卫星序列设计并合成引物22 对，其中18对引物有扩增 产物，且条带清晰，6 对引物在中国美利奴羊群内多态性丰富，平均PIC 达0.7001 ，为后期与毛品质进行 相关性分析，寻找毛用性状新型分子标记奠定了基础。 关键词：绵羊；皮肤；EST ；SSR 中图分类号：S813.3 文献标志码：A 论文编号：2009-2091 IdentificationofMicrosatellitesfromSkinofSheep Wang Zunbao, Nuerli ·Abulizi, Zhao Zongsheng, Zeng Xiancun, Wu Hongbin (CollegeofAnimalScienceandTechnology,ShiheziUniversity, Shihezi Xinjiang 832003) Abstract: 3,295 ESTs from NCBI were mined to develop EST-SSR markers. By using SSRHunter soft tools, a total of 209 SSRs were identified from 196 ESTs. The frequency of EST-SSRs was 6.3% . The dinucleotide repeat motif was the most abundant SSR, accounting for 67% , followed by 9.1% , 8.1% , 5.7% and 5.3% , respectively, for tri-, hexa-, penta- and tetra-nucleotide repeats. Among the di-nucleotide repeats, AC/TG (31%)were the most abundant type. 22 primer pairs were designed from microsatellite, 18 pairs could show clear PCR products by electrophoresis, and 6 pairs show highly polymorphism in Chinese Merino sheep. The mean PIC value was 0.7001, ranging from 0.5499 to 0.7607. These EST-SSR markers can be used for further study such as in genetic mapping, identification of quantitative trait loci (QTLs), and comparative genomics studies of sheep. Keywords:sheep; skin; EST; SSR 0 引言