Construction of a Normalized Full-Length cDNA Library of Cephalopod Amphioctopus fangsiao and Development of Microsatellite Markers  

作　　者：FENG Yanwei LIU Wenfen XU Xin YANG Jianmin WANG Weijun WEI Xiumei LIU Xiangquan SUN Guohua 

机构地区：[1]Key Laboratory of Marine Ecological Restoration, Shan Dong Marine Resource and Environment Research Institute [2]College of Fisheries and Life Science, Shanghai Ocean University [3]Key Laboratory of Marine Genetics and Breeding of Ministry of Education, Ocean University of China

出　　处：《Journal of Ocean University of China》2017年第5期897-904,共8页中国海洋大学学报（英文版）

基　　金：the National Natural Science Foundation of China (Nos. 31302215, 31272643);the Shandong Provincial Natural Science Foundation (Nos. BS2014NY010, ZR2013CQ030);the Shandong Provincial Primary Research and Development Projects (No. 2015GNC110017)

摘　　要：Amphioctopus fangsiao is one of the most economically important species and has been considered to be a candidate for aquaculture. In order to facilitate its fine-scale genetic analyses, we constructed a normalized full-length library successfully and developed a set of microsatellite markers in this study. The normalized full-length library had a storage capacity of 6.9×105 independent clones. The recombination efficiency was 95% and the average size of inserted fragments was longer than 1000 bp. A total of 3440 high quality ESTs were obtained, which were assembled into 1803 unigenes. Of these unigenes, 450(25%) were assigned into 33 Gene Ontology terms, 576(31.9%) into 153 Kyoto Encyclopedia of Genes and Genomes pathways, and 275(15.3%) into 22 Clusters of Orthologous Groups. Seventy-six polymorphic microsatellite markers were identified. The number of alleles per locus ranged from 4 to 17, and the observed and expected heterozygosities varied between 0.167 and 0.967 and between 0.326 and 0.944, respectively. Twelve loci were significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction and no linkage disequilibrium was found between different loci. This study provided not only a useful resource for the isolation of the functional genes, but also a set of informative microsatellites for the assessment of population structure and conservation genetics of A. fangsiao.Amphioctopus fangsiao is one of the most economically important species and has been considered to be a candidate for aquaculture. In order to facilitate its fine-scale genetic analyses, we constructed a normalized full-length library successfully and developed a set of microsatellite markers in this study. The normalized full-length library had a storage capacity of 6.9×105 independent clones. The recombination efficiency was 95% and the average size of inserted fragments was longer than 1000 bp. A total of 3440 high quality ESTs were obtained, which were assembled into 1803 unigenes. Of these unigenes, 450（25%） were assigned into 33 Gene Ontology terms, 576（31.9%） into 153 Kyoto Encyclopedia of Genes and Genomes pathways, and 275（15.3%） into 22 Clusters of Orthologous Groups. Seventy-six polymorphic microsatellite markers were identified. The number of alleles per locus ranged from 4 to 17, and the observed and expected heterozygosities varied between 0.167 and 0.967 and between 0.326 and 0.944, respectively. Twelve loci were significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction and no linkage disequilibrium was found between different loci. This study provided not only a useful resource for the isolation of the functional genes, but also a set of informative microsatellites for the assessment of population structure and conservation genetics of A. fangsiao.

关 键 词：microsatellite ESTs polymorphic informative normalized length alleles isolation aquaculture linkage 

分 类 号：S917.4[农业科学—水产科学]