鉴定海参的斑点杂交法及其应用  

Identification of 4 Sea Cucumber Species with Dot Hybridization and Species-Specific Probes

作　　者：律迎春[1] 段高飞[1] 左涛[1] 徐洁[1] 李晋[1] 郑荣[1] 唐庆娟[1] 薛长湖[1] 

机构地区：[1]中国海洋大学食品科学与工程学院,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2012年第11期62-65,共4页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(31101281);海洋公益性行业科研专项项目(201105029);国际科技合作项目(2010DFA31330);"泰山学者"建设工程专项(JS200510013)资助

摘　　要：本研究目的是通过斑点杂交方法对不同种类的海参进行快速准确的鉴定。采用CTAB沉淀法提取海参的DNA,PCR扩增线粒体COⅠ基因并测序,利用Primer premier 5.0软件设计并筛选了仿刺参(Apostichopus japonicus)、北大西洋瓜参(Cucumaria frondosa)、加州红参(Parastichopus californicus)及梅花参(Thelenota ananas)4种海参的特异性探针,设计并进行斑点杂交实验。实验结果显示:4条探针均具有高度的特异性,灵敏度可达100pg,能够实现对仿刺参、北大西洋瓜参、加州红参和梅花参的准确鉴定。本文建立的斑点杂交方法,为海参品种的快速、准确的鉴定提供了可靠的技术方法。Species of sea cucumber are morphologically and anatomically distinguishable with each other however such an easiness is not available for deeply processed sea cucumbers. In this study, dot hybridiza- tion method was developed in order to indentify 4 sea cucumber species （Apostichopus japonicus, Cucum- aria f rondosa , Thelenota ananas , and Parasticho pus cali f ornicus ）. Species specific probes were designed according to a 692 bp fragment of mitochondrial cytochrome I gene （CO I ） of the 4 species with Primer premier 5.0. The results showed that the probes hybridize 4 sea cucumbers, respectively, with a high specificity. The sensitivity reached 100 pg genome DNA. Dot hybridization was proved as a convenient and specific technique of identifying sea cucumber species and tracing their sources.

关 键 词：海参 线粒体COⅠ基因 斑点杂交 种类鉴定 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]