胡桃EST-SSR标记的筛选及引物设计

目的]筛选胡桃的EST-SSR分子标记并对其进行引物设计。方法]利用生物信息学方法对NCBI网上公开的5213条胡桃（Juglans regia Linn.）ESTs序列进行EST-SSR特征分析,利用利用Primer3.0软件对筛选出的EST序列设计引物。结果]在ESTs序列中共检索出207个SSR,其中无冗余序列188,检出率为3.97%,平均分布距离为21.12kb,包括92种重复基元,其中以二核苷酸和三核苷酸重复类型为主,分别占总数目的31.40%和35.27%;对随机筛选出的50条SSR引物进行PCR扩增验证,初步筛选出30对引物。结论]通过对胡桃EST序列中SSR位点的发掘分析,有针对性地设计EST-SSR引物,将为胡桃科EST-SSR分子标记的开发奠定基础。

Development of Walnut EST-SSR Markers and Primer Design

Objective] This research aimed to develop walnut EST-SSR markers and design corresponding primers.Method] 5213 EST sequences of walnut(Juglans regia Linn.) public online in NCBI were used for character analysis with bioinformatics methods,and primers were designed for the selected EST sequences by using Primer 3.0 software.Result] 207 SSRs were obtained from the EST sequences,including 188 non-redundant sequences,the detection rate was 3.97% with an average distribution distance of 21.12 kb.Totally 92 types of repeat motifs were involved,which were mainly composed of dinucleotide and trinucleotide,accounting for 31.40% and 35.27% of the total number of repeat motifs,respectively.30 pairs of primers were initially selected from the 50 randomly-selected SSR primers by PCR amplification.Conclusion] This research would lay foundations for the development of EST-SSR molecular markers in walnut and design of the targeted EST-SSR primers by mining and analyzing the SSR sites in walnut EST sequences.