海洋科学Issue(4)：1-5,5.
细角螺微卫星DNA富集文库构建及特征分析
Construction and Characterization of DNA Libraries en-riched for Microsatellites repeat sequences in Hemifusus ter-natanus
摘要
Abstract
Microsatellites enriched genome library for H. ternatanus was constructed by bio-(GT)15 and bio-(CT)15 using FIASCO. Genomic DNA was digested with restriction enzyme MseI. Fragments ranging from 300 to 1200 bp were selected to ligate with short linkers. Fragments containing microsatellite repeats were captured with Streptavidin-coated Magnetic Sphere Particles. The target fragments were eluted, PCR amplified, purified and then ligated to pMD-19T vector and transformed into Escheri- chia coli. A total of 248 clones with DNA fragments above 500 bp were selected for sequencing. As the result, 278 microsatellite sequences were isolated. Among the microsatellites, 171 repeating motifs were perfect, occupying 61.5 %; 81 repeating motifs were imperfect, occupying 29.1%; 26 repeating motifs were compound, occupying 9.4%. Besides the GT and CT repeats, several other types of repeats were also detected, such as GTT, TGG, GAA, CAA, TCTA, ACAG, TAGA, GTGA, GTCT, GATA and TTTTG. There are 82 sequences among the 278 microsatellites can be designed with primers and these primers could be used to test polymorphism in further research.关键词
细角螺(Hemifusus ternatanus)/磁珠富集/微卫星/构建/DNA文库Key words
Hemifusus ternatanus/enrichment by magnetic beads/microsatellite DNA/construction/DNA library分类
农业科技引用本文复制引用
李清荟,陈晓姣,黎中宝,曹媛钰,陈丽娜,戴刚..细角螺微卫星DNA富集文库构建及特征分析[J].海洋科学,2013,(4):1-5,5.基金项目
集美大学创新团队基金项目(2010A004) （2010A004）
