大竹蛏(Solen grandis)cDNA文库中微卫星标记的筛选  

ISOLATION AND CHARACTERIZATION OF MICROSATELLITE MARKERS FROM cDNA LIBRARY OF SOLEN GRANDIS

作　　者：乔洪金[1] 刘相全[1] 孙国华[1] 房景辉[1] 韦秀梅[1] 张滔[2] 

机构地区：[1]山东省海洋水产研究所山东省海洋生态修复重点实验室,烟台264006 [2]鲁东大学生命科学学院,烟台264025

出　　处：《海洋与湖沼》2012年第6期1128-1133,共6页Oceanologia Et Limnologia Sinica

基　　金：山东省农业良种工程项目"优质高产抗逆贝类良种选育"资助;2009-2013;水生动物营养与饲料"泰山学者"岗位经费资助;2007-2012;山东省自然科学基金(ZR2012CM037)资助;2012-2015

摘　　要：利用微卫星查找软件SSRIT对大竹蛏cDNA文库(2038条EST)中2—6个碱基重复单元组成的简单序列重复进行了筛选。最少重复次数设定为5次,共发现包含微卫星位点的EST96条,占整个EST数据库的4.71%;共发现微卫星位点103个,其中含双碱基重复序列77个,数量最多,占总数的74.76%;三、四碱基重复序列分别占微卫星序列总数的22.33%、2.91%,没有发现五或六碱基的重复。对含有SSR位点符合微卫星引物设计的EST序列,利用在线引物设计软件Primer3设计合成引物14对,以南通野生群体为模板PCR扩增和聚丙烯酰胺凝胶电泳发现,其中5对有多态性位点。在5个微卫星位点上,等位基因的数目从2—7个不等,观测杂合度和期望杂合度分别为0.067—1.000和0.066—0.775,香农指数在0.146—1.545之间。结果表明,所筛选的微卫星标记可用于遗传分析。The distribution of 2-6bp simple sequence repeats （SSRs） were analyzed from cDNA Library （2038 ESTs） of Solen grandis using the software SSRIT. When the minimum repeats were assigned as 5, 96 sequences were found to contain microsatellite sequences （taking 4.71%）. Up to 103 microsateUite loci were obtained, among which the dinucleotide repeats motifs were the most abundant type （taking 74.76%）, followed by trinucleotide and tetranucleotide repeats of 22.33%, 2.91% respectively, and pentanucleotide and hexanucleotide repeats were not found. 14 primer pairs were designed with Primer 3 software from EST sequences containing microsatellites, and 5 primers had polymorphic PCR products in Nantong wild population by PCR and polyacrylamide gel electrophoresis analysis. In these 5 microsatellite loci, the allele numbers, the observed heterozygosity and expected heterozygosity distributed from 2 to 7, 0.067 to 1.000 and 0.066 to 0.775, respectively. Shannon＇s diversity index ranged from 0.146 to 1.545. The results showed that 5 microsatellite loci were suitable for genetic analysis.

关 键 词：大竹蛏 微卫星 表达序列标签 等位基因 

分 类 号：S968.3[农业科学—水产养殖]