| 摘要: |
| 利用Vector NTI Advance 11软件分析了NCBI数据库中大菱鲆的12428条EST序列, 筛出候选SNP位点; 应用高分辨率熔解曲线(HRM)对大菱鲆不同性状群体进行基因分型。结果表明, 522个重叠群中共筛出160多个候选SNP位点; 设计56对引物用于实验, 能扩增出目的片段的引物有37条, 合45个位点, 成功率为66.1%; 设计探针, 能与候选SNP位点杂交的探针有13条, 杂交率为28.9%。本实验中能成功进行基因分型的位点共有8个, 占杂交位点的61.5%, 其中有6个为碱基替换型位点, 即G-A和C-T各占3个, 2 个为碱基颠换型位点, 即T-G和A-T各占1个。 |
| 关键词: 单核苷酸多态性标记(SNP), 大菱鲆, 高分辨率熔解曲线分析, EST 序列 |
| DOI：10.11693/hyhz201206016016 |
| 分类号: |
| 基金项目:现代农业产业技术体系专项资金(CARS-50-G01); 国家863计划(2012AA10A408-8) |
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| STUDY ON THE DEVELOPMENT OF SNP MARKERS IN SCOPHTHALMUS MAXIMUS USING HIGH-RESOLUTION MELTING (HRM) |
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LIU Qing-Ming1,2,3,4, LI Meng1,2,5,6, MA Ai-Jun1,2,3, WANG Xin-An1,2,3, HUANG Zhi-Hui1,2,3, GUO Jian-Li1,2,3
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1.Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences;2.Key Laboratory of Sustainable Development of Ma-rine Fisheries, Ministry of Agriculture;3.Qingdao Key Laboratory for MarineFish Breeding and Biotechnology;4.Shanghai Ocean Univers;5.Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology;6.Shanghai Ocean Univer
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| Abstract: |
| The experiment mines Scophthalmus maximus 12428 EST sequence from NCBI database. Putative SNP were detected from assembled contigs using Vector NTI Advance 11. SNP genotyping was performed using high-resolution melting to different character populations. Primers and probes were designed using Primer5 and Oligo7, respectively. The results show that, More than 160 putative SNP were found from 522 contigs; 56 putative SNP were selected for primer design and for SNP marker development, 37 pairs of primer were successfully amplified, including 45 putative SNP site, the success rate is 66.1%; Probes which can hybrid with Putative SNP sites have 13, hybridization Rate is 28.9%; 8 sites of hybridization sites displayed polymorphisms in different character populations, accounting for 61.5% of the hybrid sites. There are 6 transition sites, namely the G-A and C-T are 3 of each. There are 2 transversion sites, namely the T-G and A-T are 1 of each |
| Key words: Single nucleotide polymorphism (SNP), Scophthalmus maximus, High-resolution melting analysis, EST sequence |
