红鳍东方鲀BAC数据库和ESTs数据库中微卫星的筛选与应用

Identification and Application of Microsatellite Makers from BAC and ESTs Sequence in redfin puffer (Takifugu rubripes)

利用Tandem Repeats Finder软件在红鳍东方鲀Takifugu rubriper BAC数据库和ESTs数据库中查找微卫星标记。选取其中22个序列(11个BAC-SSR s序列、11个EST-SSRs序列)设计引物,并将22对引物用于庄河、河北Ⅰ和河北Ⅱ3个群体,均能扩增出产物,其中EST-SSRs有2对引物扩增产物比预期扩增产物偏大,可能是相应基因组中有内含子导致。研究结果表明,EST-SSRs标记要比BAC-SSR s标记的多态性低,其中11个BAC-SSR s和9个EST-SSRs标记有很好的多态性,可以用于红鳍东方鲀的遗传多样性分析和重要经济性状筛选等方面的研究。

Microsatellites as a new kind of molecular marker are extremely important markers, and have more advantages over the traditional genomic- derived simple sequence repeats （SSRs）. Twenty- two new microsateUites were identified from redfin puffer （ Takifugu rubripes） BAC and EST library found in UK MRC HGMP - RC （ the Fugu Genomics Project） by using bioinformatics software of Tandem Repeats Finder. Twenty - two pairs of micrsat- ellite primer were designed and applied to investigate the polymorphism in the populations of redfin puffer from Zhuanghe, Hebei I and Hebei II. All pairs of primer generated PCR amplification products, and 9 Out of 11 （ expressed sequence tags） EST - SSRs primers yielded products of expected size while 20 pairs of primer containing polymorphisms. There was lower allele number in the microsateUites located in ESTs than those isolated from a BAC library. These novel microsatellites will be very useful for population genetic and reproductive strategies studies, as well as for screening economic character of redfin puffer.