鲫鱼微卫星DNA的分离及多态性分析

Microsatellite Separation and the Polymorphism Analysis on Carassius auratus

目的:用近缘物种鲤微卫星引物来分离鲫鱼微卫星标记并对其多态性进行分析。方法:以鲫鱼基因组DNA为模板,采用6对鲤微卫星引物进行PCR扩增,PCR产物经8%的非变性聚丙烯酰胺凝胶电泳和银染色检测。结果:筛选出2个以AC和TA为重复单元的鲫鱼新的微卫星标记。多态性分析表明,这2个微卫星标记的遗传杂合度分别为0.611和0.644,多态信息含量为0.536和0.572,属于高度多态性标记。结论:该研究筛选的2个微卫星标记可应用于鲫鱼遗传多样性、遗传连锁图谱构建及分子标记辅助育种等方面的研究。

This study used the microsateUite primers of carp to isolate C, Carssius auratus L＇ s new microsatellite markers,then analyzed the polymorphism of the two microsatellite loci. Methods： 6 pairs of microsatellite primers were used to amplify the genomic DNA in Carassius auratus L using the PCR technique,The PCR products were analyzed by 8% non- denatured polyacrylamide gelatin electrophoresis, and stained with silver, then we identified the sequences of 2 pairs of primers＇s PCR products which contain microsatellite DNA fragments. Results： After the sequenees aligning and BLAST analysis , the results indicated that we isolated 2 new microsateUite sequences of Carassius auratus L, which contains the repeat units of TA and AC. Then we designed two primers according to the new microsateUite sequences and further amplified by PCR on C. auratus auratus to investigate polymorphism of the 2 microsatellite markers. The results show that the heterozygosis of the two microsatellite loci are 0.611 and 0.644,the polymorphism content are 0.536 and 0.572. Conclusion：which indicated that these two microsatellite loci have high polymorphism. The results indicated that the microsateUite markers of Carassius auratus of great importance in heredity multiplicity, heredity chain - like atlas construction, the molecular mark assistance breeding and the identify of idioplasm resources.