摘要
利用微卫星查找软件对现已公布的6632条刺参ESTs的数据库中2—6个碱基重复单元组成的简单序列重复进行筛选,进而对其微卫星的丰度和分布进行比较研究。共发现微卫星序列416个,占整个ESTs数据库的6.48%;其中含双碱基重复序列146个,数量最多,占ESTs数据库中发现微卫星序列总数的65.86%;三、四、五、六碱基重复序列分别占微卫星序列总数的26.68%、1.68%、0.24%、5.53%。对含有SSR位点符合微卫星引物设计的EST序列利用Primer软件结合人工方法设计合成引物21对,其中13对有扩增产物且均为多态位点。扩增产物变性聚丙烯酰胺胶电泳,统计每个微卫星标记等位基因数目,计算等位基因频率、观测杂合度、期望杂合度等统计学指标的评价。结果表明,在13个微卫星位点上,等位基因的数目从3到8不等,等位基因长度从175—382bp,观测杂合度和期望杂合度分别为0.158—0.650和0.198—0.841,所筛选微卫星标记可于遗传分析。
Frequency and density of 2--6bp simple sequence repeats(SSRs)were analyzed from Apostichopus japonicus EST(expressed sequence tag)database comprising 6632 sequences.The result shows that 416 microsatellite sequences(taking 6.48%)were obtained.The dinucleotide repeats motifs appeared to be the most abundant type in A.japonicus ESTs(taking 65.86%),followed by trinucleotide,tetranucleotide,pentanucleotide and hexanucleotide repeats at 26.68%,1.68%,0.24%,and 5.53%,respectively.21 primer pairs were designed with Primer software and manual-searching method from EST sequences containing microsatellites.13 primers had polymorphic PCR products in Chinese and Russian sea cucumber samples by electrophoresis analysis.In the 13 microsatellite loci,the alleles numbers distributed from 3 to 8,and the range of these alleles were between 175 to 382bp.The observed heterozygosity and expected heterozygosity distributed from 0.158--0.650 and 0.198--0.841,respectively,which showed that these 13 microsatellite loci were suitable for genetic anaivsis.
作者
孙国华
杨建敏
宋志乐
孙孝德
王卫军
张宇
SUN Guo-Hua;YANG Jian-Min;SONG Zhi-Le;SUN Xiao-De;WANG Wei-Jun;ZHANG Yu(Shandong Marine Fisheries Research Institute,Yantai,264006;Zhifu Fishery Technical Extension Station,Yantai,264001;School of Fishery and Life Science,Shanghai Ocean University,Shanghai,201306)
出处
《海洋与湖沼》
CAS
CSCD
北大核心
2010年第1期133-139,共7页
Oceanologia Et Limnologia Sinica
基金
山东省良种工程项目"速生抗病耐高温刺参良种选育":2008-2011
山东省优秀中青年科学家奖励基金(博士基金)项目:2008BS06004号
山东省科技攻关项目"海参新品系选育及构建":2008GG10005023号
关键词
刺参
微卫星
表达序列标签
等位基因
Apostichopusjaponicus
Microsatellite
Expressed sequence tag(EST)
Allele
