【作者】 彭薇； 杜慧霞； 鄢婧婧； 董颖； 周遵春； 胡景杰；

【Author】 PENG Wei1, DU Huixia1, YAN Jingjing2, DONG Ying2, ZHOU Zunchun2, HU Jingjie1 (Key Laboratory of Marine Genetics and Breeding, College of Marine Life Sciences, Ocean University of China, Qingdao 266003, Shandong, China)

【机构】 中国海洋大学海洋生命学院； 辽宁省海洋水产科学研究院；

【摘要】 采用3种不同的限制性内切酶AluI、RsaI、HaeIII,通过富集文库-菌落原位杂交法,构建了3个仿刺参微卫星DNA富集文库,共获得微卫星DNA序列583个,根据微卫星DNA的两翼序列设计了188对引物,筛选得到123个有多态性的位点。另外,对微卫星标记在仿刺参F1代家系中的分离方式进行了研究。采用45个已发表的微卫星标记,以3个F1代全同胞家系、每个家系各50个子代个体为实验材料,进行扩增分析,结果显示3个家系中偏离孟德尔分离的位点比例分别为17.39%、16.67%、19.23%。筛选仿刺参微卫星DNA标记、分析其在F1代家系中的遗传分离情况,为仿刺参微卫星遗传连锁图谱的构建、数量性状位点(QTL)的定位和选育品系的遗传分析提供了一定的理论依据。更多还原

【Abstract】 In the present study, we developed and optimized a fast and efficient method, enrichment-colony hybridization, to isolate microsatellite markers for sea cucumber (Apostichopus japonicus). A total of 3000 clones derived from the enrichment library were screened and 2167 clones gave the positive response. 800 clones were selected for sequencing and the results showed that 789 clones contain at least one microsatellite. 188 primer pairs were designed using the software Primer Premier 5.0, and the polymorphism was assessed with 48 individuals. The results showed that 123 loci were polymorphic. Also, the inheritance patterns of 45 microsatellite markers were tested in three F1 full-sib families. 23, 24 and 26 markers were polymorphic and segregated in Family One, Family Two and Family Three. 18 ratios conformed to Mendelian segregation in Family One, 20 and 21 in Family Two and Family Three.更多还原