基于mt-16S rDNA和mt-COI基因的海月水母分子生物学鉴定方法和检测技术

应用生态学报

基于mt-16S rDNA和mt-COI基因的海月水母分子生物学鉴定方法和检测技术

王建艳¹,甄毓²,王国善¹,米铁柱²,于志刚^3**

(¹中国海洋大学海洋生命学院, 山东青岛 266003; ² 中国海洋大学海洋环境与生态教育部重点实验室, 山东青岛 266100; ^3中国海洋大学海洋化学理论与工程技术教育部重点实验室, 山东青岛 266100)

出版日期:2013-03-18 发布日期:2013-03-18

Molecular identification and detection of moon jellyfish (Aurelia sp.) based on partial sequencing of mitochondrial 16S rDNA and COI.

WANG Jian-yan¹, ZHEN Yu², WANG Guo-shan¹, MI Tie-zhu², YU Zhi-gang³

(¹College of Marine Life Sciences, Ocean University of China, Qingdao 266003, Shandong, China; ^2Ministry of Education Key Laboratory of Marine Environment and Ecology, Ocean University of China, Qingdao 266100, Shandong, China; ^3Ministry of Education Key Laboratory of Marine Chemistry Theory and Technology, Ocean University of China, Qingdao 266100, Shandong, China)

Online:2013-03-18 Published:2013-03-18

摘要/Abstract

摘要：

以我国近海海域常见的海月水母为目标生物,提取其水母基因组DNA,PCR扩增mt-16S rDNA(650 bp)和mt-COI基因(709 bp)的部分序列,经纯化、克隆、测序后,将获得的序列进行BLASTn分析,筛选目的序列中与其他水母差异较大的区域,设计出8对分别针对海月水母mt-16S rDNA和mt-COI基因序列的特异性引物.经过实验室培养样品和模拟现场样品检测发现,针对mt-16S rDNA的引物AS3和mt-COI的引物AC3具有较好的特异性,可以从海蜇、沙海蜇、霞水母、端鞭水母和海月水母中快捷地检测出目标水母,从而初步建立了海月水母的分子生物学鉴定方法和检测技术.本技术摆脱了水母形态学鉴定方法存在的局限,可准确、快速地对样品进行定性分析.

Abstract: Taking the moon jellyfish Aurelia sp. commonly found in our coastal sea areas as test object, its genome DNA was extracted, the partial sequences of mt-16S rDNA (650 bp) and mt-COI (709 bp) were PCR-amplified, and, after purification, cloning, and sequencing, the sequences obtained were BLASTnanalyzed. The sequences of greater difference with those of the other jellyfish were chosen, and eight specific primers for the mt-6S rDNA and mt-COI of Aurelia sp. were designed, respectively. The specificity test indicated that the primer AS3 for the mt16S rDNA and the primer AC3 for the mtCOI were excellent in rapidly detecting the target jellyfish from Rhopilema esculentum, Nemopilema nomurai, Cyanea nozakii, Acromitus sp., and Aurelia sp., and thus, the techniques for the molecular identification and detection of moon jellyfish were preliminarily established, which could get rid of the limitations in classical morphological identification of Aurelia sp., being able to find the Aurelia sp. in the samples more quickly and accurately.

王建艳1,甄毓2,王国善1,米铁柱2,于志刚3**. 基于mt-16S rDNA和mt-COI基因的海月水母分子生物学鉴定方法和检测技术[J]. 应用生态学报.

WANG Jian-yan1, ZHEN Yu2, WANG Guo-shan1, MI Tie-zhu2, YU Zhi-gang3. Molecular identification and detection of moon jellyfish (Aurelia sp.) based on partial sequencing of mitochondrial 16S rDNA and COI.[J]. Chinese Journal of Applied Ecology.