刺参微卫星DNA分子标记的分离及筛选  

Isolation of microsatellite markers and screening microsatellite markers from sea cucumber(Apostichopus japonicus)

作　　者：丁君[1] 曹洁[1] 李润玲[1] 孙效文[1] 常亚青[1] 

机构地区：[1]大连水产学院农业部海洋水产增养殖学与生物技术重点开放实验室,辽宁大连116023

出　　处：《大连水产学院学报》2006年第2期95-99,共5页Journal of Dalian Fisheries University

基　　金：国家自然科学基金资助项目(30200212);大连市科学技术基金资助项目(990528)

摘　　要：采用磁珠富集法分离刺参Apostichopus japonicus的微卫星分子标记，共获得阳性克隆123个，其中106个含微卫星DNA序列。分析结果表明：完美型有48个，占45．28％，非完美型有39个，占36．79％，复合型有19个，占17．92％；重复碱基数以双核苷酸重复最常见，占84．91％，双核苷酸中又以CA／GT重复所占比例最大；在重复次数方面，以重复数为3-10次、11-18次和≥35次最为常见，各占39．62％、26．42％和16．98％，重复数达100次以上的有5个，最多的达到148次，有28个微卫星序列微卫星含量丰富。筛选的22对引物中，可产生出扩增产物的有17对，其中16对引物的产物带在预测区域内出现；有7对引物在中国、俄罗斯刺参模板中表现出多态性。文中还对刺参微卫星的特点进行分析，对刺参微卫星引物的PCR筛选结果进行了探讨。In the present study, microsatellites DNA was isolated and enriched from genome of sea cucumber （Apostichopusjaponicus） by magnetic beads 123 positive clones and 106 microsatellite sequence were obtained. The microsatellite sequences were structurally categorized into follows： perfect 48 （45.28%）, imperfect 39 （36. 79% ） ,and compound 19 （ 17.92% ）. Besides the CA/GT contained in the oligoprobe, CATA, CT,CCA,GA, CACT, CAA, and CACAGA repeats were found. Among the microsatellite repeats, relatively short arrays were abundant, the repeating frequence of 3 - 10, 11 - 18 and ≥35 accounted for 39.62%, 26. 42% and 16.98%, respectively. The largest array contained 148 repeats. This study will enrich microsatellite markers of sea cucumber. Base on the flanking sequence of the microsatellite loci, 22 pairs of microsatellite primer were designed and applied to investigate the polymorphism in populations of Chinese and Russian sea cucumber populations. 17 pairs of primer generated PCR amplification products, and 7 of them yield polymorphic products that can be used in the study of population genetics and genetic mapping.

关 键 词：微卫星DNA 刺参 磁珠富集 重复数 多态性 

分 类 号：S968.9[农业科学—水产养殖]