中国明对虾基因组微卫星重复单元类型与其多态性关系  

RELATIONSHIPS BETWEEN POLYMORPHISM AND TYPES OF SSR MOTIF IN THE GENOME OF CHINESE SHRIMP FENNEROPENAEUS CHINENSIS

作　　者：高焕[1] 于飞[2] 栾生[3] 孟宪红[3] 蔡生力[4] 孔杰[3] 

机构地区：[1]淮海工学院,江苏省海洋生物技术重点建设实验室,连云港222005 [2]连云港市海洋与水产科学研究所,连云港222042 [3]中国水产科学研究院黄海水产研究所,农业部海洋渔业资源可持续利用重点开放实验室,青岛266071 [4]上海海洋大学水产与生命学院,上海200090

出　　处：《水生生物学报》2009年第1期94-102,共9页Acta Hydrobiologica Sinica

基　　金：农业部“948”项目(2006-G55(B));上海水产大学农业部水产种质资源与养殖生态重点开放实验室开放课题基金(KFT2006-6);江苏省海洋生物技术重点建设实验室开放基金(2006HS004)资助

摘　　要：利用超声波粉碎中国明对虾Fenneropenaeus chinensis基因组后建立随机基因组文库,对其测序后获得了1996个克隆序列,经Seqm anⅡ(DNAstar)拼装后获得独立克隆数目为1900个,每个序列长度从400—700bp不等。利用重复序列分析软件对这些序列中含有微卫星重复序列的序列进行分析,共找到136个包含完整侧翼序列的重复序列。利用引物设计软件从以上重复序列中设计出34对引物,合成引物后,通过PCR扩增和聚丙烯酰胺凝胶电泳的方法获得了各个微卫星位点的等位基因数目。34对引物中,除4个没有扩增出产物外,其他都有较好的扩增结果,可以分辨出多态性信息情况,并据此分析了不同微卫星重复序列类型与其对应的位点多态性之间的关系。结果表明,两碱基重复类型具有较高的遗传多态性,而三碱基和四碱基以及复合型重复类型的平均多态性不高;两碱基重复序列类型各拷贝类别间的多态性信息没有明显的差异。进一步对两碱基的重复拷贝数目与多态性信息(等位基因数目)的相关关系进行分析,以考察拷贝数多少与等位基因数目之间的关系。利用SPSS软件进行相关分析,结果表明重复拷贝数目和等位基因数目呈一定相关(相关系数0.121),但相关性不显著(P=0.621)。The microsatellite （SSR） is a kind of co-dominant, and special molecular marker, and used widely in many genetics analysis, such as genetic distance between different populations of organisms. Though papers about the isolation of mierosatellite can be often found in journals, no detailed reports about which motif type of mierosatellite might be more polymorphism were found so far. Here, we wanted to isolate the microsatllite sequences from random genomic library of Chinese shrimp （F. Chinensis）, and obtained polymorphism mierosatelliie loci, then seek after the correlation between the polymorphism and the repeat motifs of mierosatellites. Using the ultrasonic to break the genome DNA, the length of about 800--1500 bp DNA were selected to establish the random clone DNA library, and 1996 sequences were sequenced using MegaBace 1000 sequencer （Amersham Bioseiences）. After the bio-soft Seqman Ⅱ （I）NAstar） assembling, 1990 in- dependent sequences were got, and the length of each clone sequence was about 400--700bp, from which 136 microsatel- lite-containing sequences were found. With the help of primer designed software （ Premier Primer 5.0） , thirty-four primer pairs were designed and used to amplify the genome of F. Chinensis by PCR. The criterion for primer design was that the repeat number of motif was more than fifteen and the length of primer sequences was between 20 and 24. The PCR amplifi- cation results were further detected by AgNO, staining methods and subsequently scanning by scanner. In these primer pairs, the detected results were as following： four primer pairs had no any amplification production, and thirty primers worked effectively, in which the polymorphism （allele） was observed. So, the success rate of primer design is 88.2%. In the 34 primer pairs, though 30 microsatellite primer pairs generated amplification products, 12 primer pairs got vivid prod- uct bands and abundant polymorphism. So, the method trying to get microsatellite primers from randomly sequenced ge- nom

关 键 词：中国明对虾 微卫星 微卫星重复单元 多态性 

分 类 号：Q343.1[生物学—遗传学]