| 应用高分辨率熔解曲线(HRM)开发大菱鲆(Scophthalmus maximus)SNP标记的研究 |
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| 引用本文： | 刘庆明,李 猛,马爱军,王新安,黄智慧,郭建丽.应用高分辨率熔解曲线(HRM)开发大菱鲆(Scophthalmus maximus)SNP标记的研究[J].海洋与湖沼,2012,43(6):1141-1148. |
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| 作者姓名： | 刘庆明 李 猛 马爱军 王新安 黄智慧 郭建丽 |
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| 作者单位： | 1. 中国水产科学研究院黄海水产研究所农业部海洋渔业可持续发展重点实验室青岛市海水鱼类种子工程与生物技术重点实验室,青岛266071 上海海洋大学,上海201306
2. 中国水产科学研究院黄海水产研究所农业部海洋渔业可持续发展重点实验室青岛市海水鱼类种子工程与生物技术重点实验室,青岛,266071 |
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| 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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| 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。
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| 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 |
| 收稿时间： | 2011/12/15 0:00:00 |
| 修稿时间： | 2012/3/21 0:00:00 |
STUDY ON THE DEVELOPMENT OF SNP MARKERS IN SCOPHTHALMUS MAXIMUS USING HIGH-RESOLUTION MELTING (HRM) |
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| LIU Qing-Ming,LI Meng,MA Ai-Jun,WANG Xin-An,HUANG Zhi-Hui and GUO Jian-Li.STUDY ON THE DEVELOPMENT OF SNP MARKERS IN SCOPHTHALMUS MAXIMUS USING HIGH-RESOLUTION MELTING (HRM)[J].Oceanologia Et Limnologia Sinica,2012,43(6):1141-1148. |
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| Authors: | LIU Qing-Ming LI Meng MA Ai-Jun WANG Xin-An HUANG Zhi-Hui and GUO Jian-Li |
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| Institution: | Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of Sustainable Development of Ma-rine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for MarineFish Breeding and Biotechnology; Shanghai Ocean Univers;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of Sustainable Development of Ma-rine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for Marine Fish Breeding and Biotechnology; Shanghai Ocean Univer;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of Sustainable Development of Ma-rine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for MarineFish Breeding and Biotechnology;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of Sustainable Development of Ma-rine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for MarineFish Breeding and Biotechnology;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of Sustainable Development of Ma-rine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for MarineFish Breeding and Biotechnology;Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Key Laboratory of Sustainable Development of Ma-rine Fisheries, Ministry of Agriculture; Qingdao Key Laboratory for MarineFish Breeding and Biotechnology |
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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 |
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| Keywords: | Single nucleotide polymorphism (SNP) Scophthalmus maximus High-resolution melting analysis EST sequence |
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