乌鳢微卫星DNA分子标记的分离及筛选

利用磁珠富集法结合放射性同位素杂交方法分离乌鳢Ophiocephalus argus的微卫星分子标记,共获得阳性克隆1 200个,从中挑选466个进行测序,得到400个含有微卫星的序列（占86.6%）。结果表明：完美型微卫星座位325个,占76.11%;非完美型微卫星座位75个,占17.57%;复合型微卫星座位27个,占6.32%。在得到的微卫星序列中,重复单元除CAG/GTC外,还观察到二碱基重复单元。根据侧翼序列设计引物139对,选择合成50对,通过优化PCR反应条件,结果有27对引物可扩增出清晰可重复的目的条带,19对引物在黑龙江群体、山东群体乌鳢的DNA样本中表现出多态性。

Isolation and screening of microsatellite markers from Argus snakehead fish Ophiocephalus argus

In this experiment,microsatellite DNA was isolated and enriched from Argus snakehead fish（Ophiocephalus argus） genome by combination of biotin capture method and radioactive labeling hybridization.About 1200 positive colonies（42.86%） were obtained through twice screens by radioactive γ 32P labeling hybridization.Sequencing of 466 positive colonies confirmed that 400 contained 427 microsatellite loci.The microsatelite sequences were structurally categorized into the following：perfect 325（76.11%）,imperfect 75（17.57%） and compound 27（6.32%）.Besides the motif of CAG/GTC contained in the colonies,there were also other dinucleotide repeats,in which 139 pairs of primers were designed with the software primer premier 3.0.In addition,50 pairs of primers were composed and screened,in which 27 pairs were used successfully to amplify special fragments,in which 19 pairs were polymorphism in the populations of Heilongjiang and Shandong Argus snakehead fish.This research offers relevant data to explo re genomes of the Argus snakehead fish.