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利用Crooijmansetal (1997)分离的包含CA重复单元的普通鲤鱼 (CyprinuscarpioL .)的 8个微卫星DNA标记 ,对从锦鲤 (CyprinuscarpioL .)的红白、大正和昭和 3个不同品系中所获得的 4个不同人工雌核发育家系的 2 0尾个体进行PCR扩增。电泳结果表明 ,8对引物在 2 0尾个体中均能重复稳定地扩增出相应的同源序列。随引物不同 ,各等位基因数为 1～ 11个 ,大小在 6 8～ 2 6 4bp。在MFW 4、MFW7、MFW 19、MFW 2 0、MFW 2 3和MFW 2 4 6个微卫星的扩增结果中 ,2 0尾个体的扩增图谱呈现了高度的遗传多态性 ,不同雌核发育家系内个体的遗传异质性也较大。其中大正 (TaS)和红白 1(RW1)的个体不仅花色分化显著 ,而且个体间的平均遗传距离分别高达 0 2 8。通过对微卫星等位基因和基因型分析发现 ,由于锦鲤品系中的每一个体是通过不断地杂交选育而获得 ,基因组来源复杂 ,基因高度杂合。因此 ,只进行 1代的人工雌核发育 ,其家系内仅部分个体的部分座位出现纯合。所获得的人工雌核发育锦鲤为后续的色素遗传调控机制研究提供了必要的实验材料 ;同时 ,所鉴定的微卫星分子标记为进行锦鲤的分子标记育种和基因组作图提供了理想的工具。
Eight microsatellite DNA markers of common carp (Cyprinus carpio L.) containing CA repeat units isolated by Crooijmansetal (1997) were used for the analysis of 4 Twenty individuals of different artificial gynogenetic families were PCR amplified. The results of electrophoresis showed that 8 pairs of primers could stably amplify the corresponding homologous sequences in 20 individuals. With the different primers, the number of alleles is 1 ~ 11, the size of 68 ~ 246bp. In the amplification results of MFW 4, MFW7, MFW 19, MFW 2 0, MFW 2 3 and MFW 2 46 microsatellites, the amplification pattern of 20 individuals showed a high degree of genetic polymorphism. Different gynogenes The genetic heterogeneity of individuals within the developmental pedigree is also greater. Among them, individuals with TaS and RW1 had significant color differentiation, and the average genetic distance between individuals was as high as 0.828. Through the microsatellite allele and genotype analysis found that because each Koi carcass strain is obtained through continuous cross breeding, the source of the genome is complex, the gene is highly heterozygous. Therefore, only one generation of artificial gynogenetic development, homozygous only part of the individual’s seat within the pedigree. The obtained artificial gynogenetic Koi provided the necessary experimental materials for the follow-up research on the genetic regulation of pigment. At the same time, the identified microsatellite markers provided an ideal tool for the molecular marker breeding and genome mapping of Koi .