一种乌鳢与斑鳢线粒体PCR-RFLP鉴定方法  

A PCR-RFLP Method for Channa argus and Channa maculate Discrimination

作　　者：董传举 张松皓 陈坤慈 宋迎楠 徐鹏[1] 孙效文[1] 

机构地区：[1]中国水产科学研究院生物技术研究中心,北京100141 [2]上海海洋大学水产与生命学院,上海201306 [3]中国水产科学院珠江水产研究所,广州510380

出　　处：《生物技术通报》2014年第8期59-64,共6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：为了高效的鉴别乌鳢与斑鳢,采用PCR-RFLP技术,对乌鳢与斑鳢开展分子生物学鉴定方法研究。通过比对乌鳢和斑鳢线粒体全序列,发现1处单核苷酸多态性位点,可以明确区分两个物种。利用1对引物对乌鳢与斑鳢线粒体基因组该区域进行PCR扩增,用限制性内切酶EcoR I分别对扩增产物进行酶切,并用1.5%的琼脂糖凝胶检测酶切结果。PCR-RFLP检测结果显示,斑鳢的PCR扩增产物被EcoR I酶切后生成两个不同大小的片段,分别为315 bp和875 bp,乌鳢则保持不变。由此可将乌鳢与斑鳢在酶切图谱上鉴别出来。In order to identificate Channa argus and C. maculate efficiently. We developed a PCR-RFLP method to conduct research in molecular biology. Briefly, two genomes of C. argus and C. maculate were aligned and compared, a SNP was identified which can discriminate two Channa species. A pair of primers was designed to amplify the mitochondrial DNA of both species, then digested by restriction enzyme EcoR I and test the digestion result by agarose gel of 1.5%. The result showed that the PCR product of C. maculate was digested by EcoR I that generate two fragments of 315 bp and 875 bp, while the PCR product of C. argus can not be digested by EcoR I. Thus, two Channa species were clearly discriminated.

关 键 词：乌鳢 斑鳢 线粒体基因组 PCR-RFLP 鉴定 

分 类 号：S917.4[农业科学—水产科学]