摘要
通过生物素磁珠富集法开发方正银鲫(Carassius auratus gibelio(Bloch))三、四核苷酸重复的微卫星,并与传统的放射性同位素杂交法相结合,进行二次筛选微卫星分子标记。结果显示:对652个阳性克隆进行测序分析,获得微卫星序列800个,其中完美型542个(67.75%),非完美型140个(17.50%),混合型118个(14.75%)。用引物设计软件primer3plus在线设计引物163对,选择合成76对,引物多态性通过二倍体野生鲫群体进行检测。检测结果显示,30对引物表现出不同程度多态性,等位基因数在3～8之间,多态信息含量(PIC)在0.4066～0.8023之间,其中80%为高度的多态性位点(PIC>0.5),表明方正银鲫微卫星文库是个高质量的文库。
Magnetic beads enriched method was used to isolated polymorphic tri-and tetranucleotide repeat microsatellite DNA from Fangzheng silver crucian carp（Carassius auratus gibelio（Bloch）） combining the traditional radioisotope hybridization.As a result,652 positive clones were sequenced after the second selection.800 microsatellite sequences were isolated from these positive clones.There were 542（67.75%） perfect motifs,140（17.50%） imperfect motifs and 118（14.75%） compound in these sequences.163 pairs of primers from these microsatellite sequences were designed with the online software primer3plus and 76 pairs of primers were synthesized among them.Results showed that 58 pairs were successful to amplify special fragments,among which 30 pairs were polymorphism in the wild crucian carp population.The number of alleles ranged from 3～8,and the PIC ranged from 0.4066～0.8023.80% loci which PIC were greater than 0.5 were in high polymorphic.The results shows that the microsatellite library of Fangzheng silver crucian carp is a library of high quality and makes a contribution to construct linkage map and analyze the genomic structure.
出处
《淡水渔业》
CSCD
北大核心
2011年第1期22-28,共7页
Freshwater Fisheries
基金
中国水产科学研究院黑龙江水产研究所基本科研专项(NO.2008HSYZX-SJ-05)
