引用本文:	王婷,黄智慧,马爱军,马得友,王新安,夏丹丹,马本贺.转录组数据的大菱鲆(Scophthalmus maximus) SNP标记开发及多态性分析.海洋与湖沼,2014,45(6):1300-1307.

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*转录组数据的大菱鲆(Scophthalmus maximus) SNP标记开发及多态性分析*
王婷,黄智慧,马爱军
中国水产科学研究院黄海水产研究所青岛市海水鱼类种子工程与生物技术重点实验室农业部海洋渔业可持续发展重点实验室,中国水产科学研究院黄海水产研究所,中国水产科学研究院黄海水产研究所

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

附件

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 (H_o) estimates ranged 0.256—1.000; expected heterozygosity (H_e) 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)