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| 基于β-连环蛋白基因的分子标记甄别海蜇(Rhopilema esculentum)和沙海蜇(Nemopilema nomurai) |
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杨傲傲1,2, 朱玲2, 周春娅2, 杨洪1,2, 骆晓蕊1,2, 刘春胜2, 庄志猛2
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1.上海海洋大学水产与生命学院 上海 201306;2.农业部海洋渔业可持续发展重点实验室 山东省渔业资源与生态环境重点实验室 中国水产科学研究院黄海水产研究所 青岛 266071
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| 摘要: |
| 采用转录组454 GS FLX 测序和PCR 技术, 以海蜇(Rhopilema esculentum)和沙海蜇(Nemopilema nomurai)的基因组DNA 为模板, 分别克隆了包含EST-SSR 和EST-SNP 标记的 β-连环蛋白基因目的片段。生物信息学分析显示, 海蜇和沙海蜇的β-连环蛋白基因目的片段长度分别为166/169 bp 和157/160 bp, 均没有内含子。海蜇个体之间β-连环蛋白基因目的片段除了微卫星重复差异外, 只有一个碱基的差异; 沙海蜇个体之间除了微卫星重复差异外, 其余完全一致。在海蜇和沙海蜇β-连环蛋白基因目的片段的相同位置均包含微卫星重复, 但其重复单元截然不同: 海蜇为:(TGC)4-6(TGT)1-2(TGC)4-5, 而海蜇为(TGT)5-6。同时两物种间还存在14 个单核苷酸多态位点: (T/C)1,(T/C)2, (C/T)3, (C/T)4, (C/T)5, (T/G)6, (G/C)7, (T/G)8, (A/G)9, (C/T)10, (G/A)11, (A/G)12, (C/T)13, (A/T)14。聚丙烯酰胺凝胶电泳图谱也直接客观地反映了两个群体之间目的片段的长度和微卫星多态性差异。上述结果显示, EST-SSR 和EST-SNP 标记的β-连环蛋白基因目的片段, 可以作为一种简单、有效的分子标记, 快速、准确地识别不同发育阶段的海蜇和沙海蜇。 |
| 关键词: 海蜇 沙海蜇 β-连环蛋白 EST-微卫星 EST-单核苷酸多态位点 |
| DOI：10.11693/hyhz20140500136 |
| 分类号: |
| 基金项目:国家重点基础研究发展计划(973)项目资助, 2011CB403605 号; 国家自然科学基金资助项目, 31372507 号; 科技基础性工作专项, 2013FY110700 号。 |
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| USING β-CATENIN GENE TO DIFFERENTIATE RHOPILEMA ESCULENTUM FROM NEMOPILEMA NOMURAI |
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YANG Ao-Ao1,2, ZHU Ling2, ZHOU Chun-Ya2, YANG Hong1,2, LUO Xiao-Rui1,2, LIU Chun-Sheng2, ZHUANG Zhi-Meng2
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1.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;2.Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Shandong Provincial Key Laboratory of Fishery Resources and Eco-environment, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
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| Abstract: |
| The target fragments of β-catenin gene including EST-SSR and EST-SNP markers were cloned with genomic DNA templates of Rhopilema esculentum and Nemopilema nomurai by the 454 GS-FLX transcriptome sequencing and PCR techniques, respectively. Bioinformatic analysis showed that target fragments nucleotide sequences did not contain intron and the lengths of nucleotide sequences were 166/169bp and 157/160bp in R. esculentumi and N. nomurai populations, respectively. There was only a base difference among individuals of R. esculentumi, and no difference among individuals of N. nomurai, apart from SSR difference. In the same site, the target fragments of β-catenin gene shared a distinct repeat motif, (TGC)4-6(TGT)1-2(TGC)4-5 in R. esculentumi and (TGT)5-6 in N. nomurai. In addition, 14 SNP loci from the target fragments of β-catenin gene were detected in two populations, including (T/C)1, (T/C)2, (C/T)3, (C/T)4, (C/T)5, (T/G)6, (G/C)7, (T/G)8, (A/G)9, (C/T)10, (G/A)11, (A/G)12, (C/T)13, and (A/T)14. SSR profile from β-catienin also reflected clearly the differences in the length and polymorphism of the target fragment of β-catenin among individuals of two populations. Therefore, the target fragments of β-catenin gene including EST-SSR and EST-SNP markers, as a simple effective molecular marker, can quickly identify R. esculentumi and N. nomurai at different developmental stages. |
| Key words: Rhopilema esculentum Nemopilema nomurai β-catenin EST-microsatellite EST-single nucleotide polymorphism |
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