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利用磁珠富集法,构建了缢蛏(Sinonovacula constricta)微卫星富集文库。开发并筛选出18对微卫星引物,其中8对微卫星引物亲缘排除率较高,选取这8对引物对家系进行了亲子鉴定。结果表明,18对微卫星引物在12个个体中等位基因为2~6,平均等位基因为3.00;观测杂合度在0~1.000之间,平均值为0.581 3;期望杂合度0.290~0.779,平均值为0.528 4;多态信息含量为0.239~0.703,平均值为0.432 2。当两个亲本基因型未知时,8对微卫星引物在整个家系中单个亲本排除率(E-1P)为0.583~0.916,平均值为0.790 0;当一个亲本基因型已知时,另一个亲本的排除率(E-2P)为0.406~0.830,平均值为0.653 9。同时可知在子代个体数量不断增加的情况下,排除率仍在较高范围,因此可以作为亲子鉴定的分子标记。这对于缢蛏规范化养殖、家系选育的开展具有重要意义。
Using enrichment of magnetic beads, a microsatellite-enriched library of Sinonovacula constricta was constructed. 18 pairs of microsatellite primers were developed and screened. Among them, 8 pairs of microsatellite primers had high rejection rate. Eight pairs of primers were selected for paternity testing of pedigrees. The results showed that the alleles of 18 pairs of microsatellite primers were 2 ~ 6 in 12 individuals and the average allele was 3.00. The observed heterozygosity ranged from 0 to 1.000 with the average value of 0.581 3. The expected heterozygosity ranged from 0.290 to 0.779. The average value was 0.528 4; the polymorphism information content was 0.239 ~ 0.703, the average was 0.432 2. When the two parents were unknown genotypes, the single-parent exclusion rate (E-1P) of 8 pairs of microsatellite primers in the entire pedigree ranged from 0.583 to 0.916 with an average of 0.790 0. When one parent genotype was known, the other parent The exclusion rate (E-2P) was 0.406-0.830 with an average of 0.653 9. At the same time we can see that in the case of increasing number of offspring individuals, the exclusion rate is still high, so it can be used as a paternity test molecular marker. This is of great significance for the development of standardized breeding and pedigree breeding.