四种鲟鱼线粒体PCR-RFLP鉴定方法的研究  

PCR-RFLP Analysis of mt DNA to Identify Four Kinds of Sturgeons

作　　者：董传举 刘园园[2] 刘晓勇[3] 宋迎楠 徐鹏[2] 孙效文[2] 

机构地区：[1]上海海洋大学水产与生命学院,上海201306 [2]中国水产科学研究院水产应用基因组中心,北京100141 [3]中国水产科学研究院鲟鱼国家级良种场,北京100070

出　　处：《生物技术通报》2014年第12期78-85,共8页Biotechnology Bulletin

基　　金：国家"863"计划项目(2011AA100401);农业部公益性行业科研专项项目(200903045)

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TaqαI、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。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 Taqa I, Ava If, Eag 1-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.

关 键 词：鲟鱼 线粒体DNA PCR-RFLP 鉴定方法 分子标记 

分 类 号：S917.4[农业科学—水产科学]