诸氏鲻虾虎鱼多态性微卫星标记的开发及评价

目的 开发诸氏鲻虾虎鱼（Mugilogobius chulae）多态性微卫星标记,为诸氏鲻虾虎鱼的遗传背景评价提供基础数据。方法 采用磁珠富集法,以生物素标记的(AC)15为探针,构建了诸氏鲻虾虎鱼微卫星富集文库。经克隆、筛选、测序后,设计合成微卫星引物,利用野生群体对微卫星引物进行多态性筛选和评价。结果 共获得74个含有微卫星重复单元的序列,根据微卫星序列共设计出33对微卫星引物。利用30个野生诸氏鲻虾虎鱼样本对33对引物的微卫星位点进行了评价,其中有12个位点表现出多态性,平均有效等位基因数（Ne）、观测杂合度（Ho）、期望杂合度（He）及多态信息含量（PIC）分别为2.9167、2.2311、0.4583、0.5633和0.4474。结论 本研究为我国本土小型海水鱼类诸氏鲻虾虎鱼微卫星标记的首次开发,所筛选的微卫星标记将为诸氏鲻虾虎鱼遗传质量控制及监测提供了实验依据。

Isolation and characterization of polymorphic microsatellite markers in Mugilogobius chulae

Objective To isolate microsatellite DNA markers from genome of Mugilogobius chulae for further identification of its genetic background. Methods A microsatellite-enriched genomic library for Mugilogobius chulae was constructed by magnetic-bead enrichment method using biotinylated (AC)₁₅ probe. After a series of steps, the obtained microsatellite markers were assessed and evaluated versus that of wild population of Mugilogobius chulae. Results 74 repeated sequences containing microsatellite sequences were obtained from 95 randomly picked positive clones. 33 primers were designed and synthesized. All of these loci were selected for polymorphism analysis in a wild population of Mugilogobius chulae, and 12 of them showed polymorphism. The mean number of alleles (Na), the number of effective alleles (Ne), observed heterozygosity (Ho), expected heterozygosity (He), and polymorphic information content (PIC) were 2.9167, 2.2311, 0.4583, 0.5633 and 0.4474, respectively. Conclusions The microsatellites developed in this study represent the first nuclear markers described in Mugilogobius chulae and will be useful tools for genetic studies of Mugilogobius chulae.