四种鲟鱼线粒体 PCR-RFLP 鉴定方法的研究

doi:10.13560/j.cnki.biotech.bull.1985.2014.12.013

摘要/Abstract

摘要： 利用线粒体序列开发高效准确的分子鉴定方法广泛应用于水产品易混物种的鉴定中。应用 PCR-RFLP 技术，对鲟鱼易混种开展了分子生物学鉴定方法研究。结果表明，利用一组引物对 8 种鲟鱼线粒体基因进行 PCR 扩增，分别应用限制性内切酶 Taqα I、Ava II 和 Eag I-HF、Nae I 对扩增产物进行酶切，并用 3.0% 的琼脂糖凝胶检测 PCR 产物的酶切结果，可从 8 种鲟鱼易混种中分别鉴别出中华鲟、小体鲟、达氏鳇以及欧鳇。所建立的方法操作简单，在保证鱼种存活基础上只需剪取少量鳍条，便可快速准确地进行常见鲟鱼和不同鲟鱼产品的鉴别，大大增加了鉴定结果的准确性和可信度，极大地提高了工作效率。

关键词: 鲟鱼, 线粒体, DNA, PCR-RFLP, 鉴定方法, 分子标记

Abstract: With the high efficient and accurate, molecular identification methods are widely used in some easily confused species. So we carried out the research in some miscible species of sturgeons with the application of PCR-RFLP. With the help of one pair primers, we amplified one region of mtDNA in 8 kinds of sturgeons’which contain Acipenseridae sinensis, A. schrenckii, A. baeri, A. ruthenus, Huso dauricus, H. huso, A. stellatus, A. gueldenstaeti. When digested with the restriction enzyme of Taqα I, Ava II, Eag I-HF and Nae I then test the results of PCR and enzyme digestion reactions with the density 3.0% of agarose gel, A. sinensis, A. ruthenus, H. dauricus, H. huso can be identified from other sturgeons. This method just need one pair of primers which is specific for binding to the mitochondrial DNA to amplify genomic DNA, then digested the products of PCR with different enzymes. So it is convenient to operate and can identify the species accurately in a relatively short period of time. On the basis of a little fin, we can identify common sturgeon species credibility and accuracy and also greatly improved the work efficiency.

Key words: Sturgeon, mtDNA, PCR-RFLP, Identification method, Molecular marker

董传举, 刘园园 ,刘晓勇3 ,宋迎楠, 徐鹏 ,孙效文. 四种鲟鱼线粒体 PCR-RFLP 鉴定方法的研究[J]. 生物技术通报, 2014, 0(12): 78-85.

Dong Chuanju, Liu Yuanyuan, Liu Xiaoyong,Song Yingnan,Xu Peng,Sun Xiaowen. PCR-RFLP Analysis of mtDNA to Identify Four Kinds of Sturgeons[J]. Biotechnology Bulletin, 2014, 0(12): 78-85.

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