文冠果富集SSR文库的构建及特性分析  

Construction and characterization of enriched microsatellite library from Xanthoceras sorbifolia

作　　者：程月琴[1] 王红卫[1] 万惠霞[1] 段景勉[1] 张佩[1] 韩卫丽[1] 

机构地区：[1]河南农业大学植物保护学院,河南郑州450002

出　　处：《河南农业大学学报》2013年第4期440-445,共6页Journal of Henan Agricultural University

基　　金：国家自然科学基金项目(31170351);河南省教育厅自然科学基金项目(2008A180012)

摘　　要：应用Dynal磁珠-生物素标记的微卫星探针(AC)8,(AG)8和(ATG)12与文冠果基因组DNA酶切片段杂交,捕获含有微卫星序列的DNA片段,连接到pMD18-T载体上,转入感受态细胞Trans5α,构建文冠果富集微卫星文库.利用M13F和M13R载体序列引物筛选文库,对插入片段长度为400～1500bp的克隆进行测序.共获得123条序列,55条(44.7%)含有微卫星位点,其中完美型占63.7%,非完美型22.4%,混合型10.3%.微卫星重复基元中,二核苷酸(AC)n最为常见.The DNA fractions containing microsatellite sequences were captured by hybridizating the digested Xanthoceras sorbifolia genomic DNA fragments with the oligonucleotide probes （AC） s, （AG） s and （ATG）12 attached to striptavadin coated magnetic beads （Dynal）. The enriched DNA fragments were ligated into pMD18-T easy vector and then transformed into competent TransSα cell to form an en- riched microsatellite sequence library. PCR screening using MI3 Forward and M13 Reverse as primers identified 123 clones containing 400 to 1 500 bp inserts. Sequence analysis of these positive clones confirmed 55 microsateUite sequences, with an enrichment efficiency of 44.7%. Of these microsatel- lite loci, 63.7% were perfect repeat motif, 22.4% imperfect and 10.3 compound repeat motifs. A- mong them, （AC）, was the most common motif.

关 键 词：文冠果 微卫星 磁珠 富集文库 

分 类 号：Q949.9[生物学—植物学]