许氏平鲉微卫星标记的开发及评价

Isolation and characterization of microsatellite markers for black rockfish Sebastes schlegeli

采用磁珠富集法筛选具多态性的许氏平鲉微卫星标记,并用一个野生群体对其多态性进行了评价。三引物PCR法筛选出58个阳性克隆进行了测序,共得到47个含微卫星核心的DNA序列,筛选出15对具有多态性的微卫星引物。所有位点上共得到66个等位基因片段,每个位点的观测等位基因数（A）为2~14个,平均等位基因数为4.4,观测杂合度（Ho）、期望杂合度（He）范围分别为0.062 5~0.968 8、0.061 5~0.929 1。每个位点的多态信息含量（PIC）显示,有6个位点表现出高度或中度多态。经Bonferroni校正后,HJ2-32、HJ4-27、HJ4-45和HJ4-47四个位点的等位基因频率显著偏离了Hardy-Weinberg平衡。连锁不平衡检测表明,各位点间无连锁不平衡现象。

and Environment Research Institute; Polymorphic microsatellite markers were developed from black rockfish（ Sebastes schlegelii） with an enriched partial genomic library by magnetic beads and the polymorphisms of these loci were assessed in 32 individuals from a wild population. We screened 58 positive clones by three-primers PCR method and sequenced them,then 47 sequences were detected containing microsatellites,and 15 pairs of polymorphic microsatellite primers were obtained. A total of 66 alleles were detected,with the number of alleles ranging from 2 to 14（ averaging 4. 4）. The observed and expected heterozygosity values ranged from 0. 062 5 to 0. 968 8 and 0. 061 5 to 0. 929 1,respectively. The values of polymorphism information content indicated,six loci were highly or moderately polymorphic. HJ2-32,HJ4-27,HJ4-45 and HJ4-47 significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction and no significant linkage disequilibrium was found. The result demonstrates these markers will facilitate relevant research in S. schlegelii.