| 摘要: |
| 大菱鲆(Scophthalmus maximus)是最具商业价值和养殖前途的海水鱼类之一, 雌雄间生长有一定的差异。其高通量雌雄转录组测序的完成为大规模鉴定和开发SNP标记提供了参考序列。本研究基于大菱鲆雌雄转录组测序数据, 选择其中45个SNP位点, 设计引物63组, 其中21个位点(46.7%)应用小片段高分辨率熔解曲线(HRM)技术分型成功。对其进行多态性检测, 21个位点均具有二个单倍型。观测杂合度Ho的分布范围为0.256—1.000, 期望杂合度He的范围为0.276—0.518, 19个位点符合Hardy-Weinberg平衡。14个SNP位点位于基因编码区, 其中3个属于非同义突变。含有这些SNP位点的基因大多与信号转导和转录翻译相关。本研究结果表明, 高通量转录组测序和小片段HRM适合大规模SNP标记开发, 为大菱鲆的分子遗传育种提供了候选标记资源。 |
| 关键词: 大菱鲆 单核苷酸多态性(SNP) 转录组 高分辨率溶解曲线(HRM) |
| DOI：10.11693/hyhz20140400101 |
| 分类号: |
| 基金项目:国家高技术研究发展计划(863)项目资助, 2012AA10A408-8 号; 现代农业产业技术体系建设专项资金资助, CARS-50-01 号 |
|
| DEVELOPMENT AND POLYMORPHIC ANALYSIS OF SNP MARKERS IN SCOPHTHALMUS MAXIMUS BASED ON TRANSCRIPTOME DATABASE |
|
Wang Ting,Huang Zhi-Hui and Ma Ai-Jun
|
|
Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences,Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences,Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences
|
| Abstract: |
| Turbot Scophthalmus maximus is a commercially valuable fish and one of the most promising aquaculture species. Completion of turbot high-throughput transcriptome sequencing provides a reference sequence to identify and develop large-scale SNP markers. Based on Scophthalmus maximus transcriptome sequencing data, we selected 45 SNPs and designed 63 pairs of primers, of which 21 (46.7%) loci using high-resolution melting (HRM) technology with small amplicons genotyping success. All the 21 sites have two haplotypes, and proved polymorphic. The observed heterozygosity (Ho) estimates ranged 0.256—1.000; expected heterozygosity (He) estimates 0.276—0.518, and 19 loci were in Hardy- Weinberg equilibrium after Bonferroni correction. Fourteen SNPs were located in the coding region, 3 non-synonymous changes. Most SNP-containing genes were related to signal transduction mechanisms and transcription. This study shows that high throughput transcriptome sequencing and small amplicons of HRM is suitable for large-scale SNP marker development, which provides a number of candidate markers resources to molecular genetic breeding of turbot. |
| Key words: turbot Scophthalmus maximus single nucleotide polymorphisms (SNP) transcriptome high resolution melting (HRM) |
