基于EST数据的柞蚕SSR标记开发

Development of SSR Markers Based on Expressed Sequence Tags in Chinese Oak Silkmoth,Antheraea pernyi

柞蚕SSR标记的开发对研究柞蚕的遗传与进化、分子遗传图谱构建、功能基因定位和克隆等有重要意义。从1 500条柞蚕表达序列标签（EST）中鉴定了71个SSR位点,平均5.2 kb出现1个SSR位点,在1~6个碱基的重复基元中,以3个和4个核苷酸重复为主导类型。设计合成了29对EST-SSR引物,通过PCR扩增和电泳检测鉴定了7对具有多态性的引物,测序验证结果表明其中的4对为有应用价值的EST-SSR标记。建立的利用EST数据开发柞蚕SSR标记的方法,有助于进一步大量开发柞蚕SSR分子标记。

Development of SSR markers in Chinese oak silkworm（ Antheraea pernyi） plays an important role in studies on genetics and evolution,construction of molecular genetic map,positional mapping and cloning of functional genes. In this study,71 simple sequence repeats（ SSRs） loci were identified in 1 500 expressed sequence tags（ ESTs） of A. pernyi. On an average,one SSR was found per 5. 2 kb of EST sequence. Among mono- to hexa-nucleotide repeat motifs,tri-nucleotide and tetra-nucleotide repeats are the dominant motifs. Twenty-nine pairs of EST-SSR primers were designed and synthesized,and 7 pairs of polymorphic primers were identified through PCR amplification and electrophoresis detection. After further sequencing,4 EST-SSR markers are considered to have application value. The established method will contribute to development of SSR molecular markers based on A. pernyi EST database in a large scale.