克氏原螯虾微卫星文库构建及多态性分析  

Construction and Polymorphism Analysis of a Microsatellite Library in Red Swamp Crayfish Procambarus clarkii

作　　者：宋丹丹 王龙 魏红静 史文竞 朱传坤 SONG Dandan;WANG Long;WEI Hongjing;SHI Wenjing;ZHU Chuankun(Jiangsu Engineering Laboratory for Breeding of Special Aquatic Organisms,School of Life Science,Jiangsu Collaborative Innovation Center of Regional Modern Agriculture&Environmental Protection,Huaiyin Normal University,Huai an 223300,China)

机构地区：[1]淮阴师范学院生命科学学院江苏省特色水产繁育工程实验室江苏省区域现代农业与环境保护协同创新中心

出　　处：《水产科学》2019年第6期819-826,共8页Fisheries Science

基　　金：江苏高校品牌专业建设工程资助项目(TAPP);江苏省大学生创新训练计划校级实践创新项目(201619008XJ)

摘　　要：采用磁珠富集法构建了克氏原螯虾(GT)n重复类型的微卫星富集文库。随机检测了82个克隆,其中有78个为阳性,阳性率达95.1%。对75个阳性单克隆进行测序后,共获得序列57条,其中43条含有微卫星序列,微卫星富集效率为75.4%。根据43条含微卫星的序列共设计了21对引物,其中18对可扩增出稳定条带,并且有9对表现为多态性。在测试群体中对9个多态标记的检测结果显示,等位基因数为2~6个(平均4个),观测杂合度为0.3522~0.6667(平均0.5856),期望杂合度为0.3704~0.8390(平均0.6291),多态信息含量为0.2859~0.8001(平均0.5584)。本研究开发的微卫星标记将为今后克氏原螯虾及其近缘物种的群体遗传结构分析、种质资源保护、遗传图谱构建等遗传学和基因组学研究提供有利工具。In this study,a microsatellite library with(GT)n repeats was constructed for red swamp crayfish Procambarus clarkii using the magnetic bead enrichment method.A total of 82 clones were sequenced randomly,78 clones of which were positive with a positive rate of 95.1%.Among 75 positive single clones,57 sequences were successfully sequenced,in which 43 clones contained microsatellites with an enrichment efficiency of 75.4%.Twenty-one pairs of primers were successfully designed for the 43 microsatellite-containing sequences with 9 pairs being polymorphic.The polymorphism tests revealed that the number of alleles was ranged from 2 to 6(4 alleles per locus averagely)in a red swamp crayfish population for these nine markers;observed heterozygosity was ranged from 0.3522 to 0.6667(0.5856 on average);expected heterozygosity ranged from 0.3704 to 0.8390(0.6291 on average);polymorphic information content ranged from 0.2859 to 0.8001(0.5584 on average).The microsatellite markers developed in this study will provide useful tools for future research including genetic mapping,analysis of genetic diversity,and molecular breeding of red swamp crayfish and close-related species.

关 键 词：克氏原螯虾 磁珠富集法 微卫星文库 多态性 

分 类 号：Q785[生物学—分子生物学]