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采用磁珠富集法(Fast isolation by AFLP sequences containing repeats,FIASCO)构建“光合1号”新品种中华绒螯蟹Eriocheir sinensis基因组微卫星文库。用PCR法对225个单菌落进行鉴定,共获得168个阳性克隆,测序发现有94个含有微卫星座位,其准确率为55.95%,其中单元重复次数为5～10的序列占34.04%,重复次数为10～20的序列占20.21%,重复次数大于20的序列占45.75%,重复次数最高达到92次;完美型微卫星座位69个(占73.40%),非完美型微卫星座位17个(占18.09%),混合型微卫星座位8个(占8.51%);根据微卫星侧翼序列设计并合成53对引物,检测结果显示,有31对(占58.50%)引物能够扩增出特异性条带;选择其中14对多态性良好的引物用于中华绒螯蟹辽河野生群体的遗传多样性分析,共检测到148个等位基因,单个位点等位基因数为7～13个,平均为10.6个,观测杂合度为0.214 3～0.666 7,期望杂合度为0.825 4～0.910 1,多态性信息含量为0.766 3～0.865 4。
The microsatellite DNA library of Eriocheir sinensis genome was cloned by Fast isolation by AFLP sequences containing repeats (FIASCO). A total of 225 positive clones were identified by PCR. A total of 168 positive clones were obtained. A total of 94 microsatellite loci were found, with an accuracy rate of 55.95%. Sequences with 5 to 10 repeat units accounted for 34.04% 20.21% of which were sequences of 10-20%, 45.75% of sequences whose repetition number was more than 20 and 92 times of repetition, 69 microsatellites (73.40%), 17 microsatellite loci Accounting for 18.09%). There were 8 hybrid microsatellite loci (accounting for 8.51%). According to the microsatellite flanking sequences, 53 pairs of primers were designed and synthesized. 31 pairs (58.50%) of the primers could amplify the specific bands Among them, 148 primers were selected for the genetic diversity analysis of Liaohe population of E. sinensis. A total of 148 alleles were detected. The number of alleles per locus ranged from 7 to 13, with an average of 10.6, the observed heterozygosity was 0.214 3 ~ 0.666 7, the expected heterozygosity was 0.825 4 ~ 0.910 1, and the polymorphism information content was 0.766 3 ~ 0.865 4.