藏羚羊基因组微卫星特征分析与引物验证-生态学专业论文.docxVIP

万方数据 万方数据 藏羚羊基因组微卫星特征分析与引物验证 中文摘要 藏羚羊全基因组序列公布后，充分研究和获取基因组微卫星序列的数据特征以及从全 基因组序列中查找具有多态性的微卫星座位，开发准确高效的微卫星引物，对藏羚羊及近 缘物种的保护遗传学研究具有重要意义。 本研究以藏羚羊基因组数据库为基础，利用 MISA 软件对藏羚羊基因组进行微卫星分 离，分析研究藏羚羊核基因组 DNA 中微卫星分布情况和特征。同时结合引物设计软件 Primer3.0 自动设计生成微卫星引物，设定相关随机抽取的参考标准，抽取部分引物，通 过聚丙烯酰胺凝胶电泳检测筛选具有多态性的微卫星位点，进而通过毛细管凝胶电泳检测 验证引物多态性是否一致。 研究结果如下： 1、以 2696.89Mb 的藏羚羊全基因组数据库为基础，共查找到 723135 个微卫星座位， 其中完全型微卫星有 675809 个。 2、在六种微卫星重复类型中，单核苷酸有 471142 个占 69.72%，是比重最多的一类 微卫星；其次是二核苷酸、三核苷酸，分别为 88832 个和 86658 个，占总数的 13.14%和 12.82%；六核苷酸含量最少，仅 215 个，约占 0.03%。其中，单核苷酸、四核苷酸、五核 苷酸以及六核苷酸中 A/T 重复单元比较丰富；二核苷酸重复类型中，最常见的是 AC/GT 重复单元，其次是 AT/TA 重复单元，而 GC/CG 重复单元数量最少；三核苷酸中，AGC/GTC 数量最多，其次是 ACG/CTG 和 AAC/GTT，而 CCG/CGG 数量最少。 3、从自动设计的 46286 对微卫星引物，随机抽选 100 对藏羚羊微卫星引物，共获得 8 对具有多态性的微卫星引物。 关键词：藏羚羊，基因组，微卫星，验证 I Genome-Wide Characterization and Verification of Simple Sequence Repeats in the Chirus Abstract After the whole genome sequence of Tibetan antelope published, it’s necessary and meaningful to study and gain the data characteristic about the microsatellite sequences of the whole genome and search the microsatellite locis with polymorphism. The study analysed the microsatellite distribution and characteristics in the nuclear genome DNA on the basis of the whole genome database .Combined MISA software and using the primers designing software( Primer 3.0), randomly selected some primers according to the set of standards.By polyacrylamide gel electrophoresis and capillary gel electrophoresis detection to verify whether the results about polymorphism were consistent. The main results were as follows: To investigate the distribution and feature of simple sequence repeats (SSRs) or microsatellites (1-6 bp repeats), we screened the entire chiru genome (2696.89 Mb) by using MISA. A total of 723135 SSR loci were detected and characterized within the whole genome, which included 675809 perfect SSRs. Among the six types of SSRs, the most abundant repeat type was mononucleotide (69.72%), followed by di- (13.14%), tri- (12.82%), penta- (2.2