卵形鲳鲹微卫星分子标记的筛选

卵形鲳鲹微卫星分子标记的筛选 Screening of Microsatellite Molecular Marker in Trachinotus ovatus 陈秀荔 CHEN Xiuli 1 first-author 肖群平 XIAO Qunping 1 陈晓汉 CHEN Xiaohan 1 彭敏 PENG Min 1 李咏梅 LI Yongmei 1 广西水产研究所/广西水产动物遗传育种与健康养殖重点实验室 广西水产研究所/广西水产动物遗传育种与健康养殖重点实验室 Fisheries Research Institute of Guangxi/Key Laboratory of Aquatic Genetic Breeding and Health Cultivation of Guangxi Fisheries Research Institute of Guangxi/Key Laboratory of Aquatic Genetic Breeding and Health Cultivation of Guangxi 本研究通过生物素与链霉素的强亲和性原理,采用生物素-磁珠吸附微卫星富集法筛选卵形鲳鲹的微卫星分子标记序列.从201个菌落中挑选出152个阳性克隆进行测序,成功测序137个,微卫星含量达到90.13%,共得到131个重复次数在6次以上的微卫星DNA序列,除生物素探针中使用的CA重复外,还得到TG、TC、GA、ATTT的重复序列.其中完美型85个,占64.9%;非完美型36个,占27.5%;混合型10个,占7.6%.利用PrimerPremier5.0设计引物90对,挑选其中的60对引物进行合成和PCR筛选,结果显示,35对引物可扩增出特异性条带.筛选的卵形鲳鲹微卫星分子标记具有多态性,可用于遗传多样性分析. Based on the theory of strong affinity principle of biotin and streptomycin,the method of bio-bead adsorption and enriched microsatellite sequence are adopted to screen Trachinotusovatusmicrosatellite markers sequence in this study.152positive clones from 201colony were sequenced,and 137of them were successfully sequenced,while the contents of microsatellite is up to 90.13%.There are a total of 131microsatellite DNA sequences which number of repetition is 6times or more.Besides the biotin-labeled SRS probes CA,TG,TC,GA and ATTT were included in the microsatellite sequence too.Among the microsatellite sequence,85repeat motifs were perfect（about 64.9%）;36repeat motifs were imperfect（about 27.5%）;10repeat motifs（about 7.6%）were compound.90pairs of primers were designed with the software Primer Premier 5.0,and 60pairs of primers were composed and screened by PCR.The results showed that 35pairs of primers can amplify special fragments.Screened microsatellite molecular marker in Trachinotus ovatus possess polymorphism,which can be used to analyze genetic diversity. 卵形鲳鲹 磁珠富集法 微卫星 Trachinotus ovatus enrichment by magnetic beads microsatellite 10.14188/j.1671-8836.2010.05.017 S917.4 国家科技支撑计划项目（2007BAD29B03） 2010-05-01 2021-04-01 5 56