利用PCR-RFLP技术对鲫鱼6个不同品系的鉴定  

Identifying Six Strains of Carassius auratus by PCR-RFLP on mtDNA ND3/ND4L/ND4 Gene

作　　者：庄怡[1] 刘必谦[1] 唐杰[1] 

机构地区：[1]宁波大学应用海洋生物技术教育部重点实验室,宁波315211

出　　处：《生物技术通报》2012年第11期144-149,共6页Biotechnology Bulletin

基　　金：浙江省海洋与渔业局项目(HK11002)

摘　　要：采用PCR-RFLP方法分析鲫鱼不同品系mtDNA ND3/ND4L/ND4基因片段,对野鲫、异育银鲫、红鲫、白鲫、彩鲫及金鱼的基因片段进行测序,测序结果用RESearch-version1.0软件对限制性内切酶特异性位点进行分析,最终选择MvaⅠ核酸限制性内切酶对目的基因片段进行酶切,同时引入鲤鱼作为外类群,构建了系统树。酶切结果分析表明,基于聚合酶链式反应——限制性片段长度多态性分析能快速而准确地区分鲫鱼的4个不同品系,首次提出异育银鲫和浙江地区野鲫(河鲫鱼)在mtDNAND3/ND4L/ND4基因上的分子鉴定法,为物种鉴定提供了一定的理论研究基础。但野鲫、金鱼、彩鲫的基因序列两两只有一个碱基的差异,相似度接近100%,用酶切软件(RESearch-version1.0)进行分析后未能找到区别三者的限制性内切酶位点,这一结果也验证了关于金鱼、彩鲫起源于野鲫的观点。Different strains of mtDNA ND3/ND4L/ND4 gene fragments for different Carassius auratus were analyzed by means of PCRRFLP,in which the gene fragments of C.auratus auratus,C.auratus gibelio,C.auratus red var,C.auratus cuvien,C.aurtus color variety and C.auratus auratus goldfish were sequenced.The sequence results were analyzed by RESearch-version 1.0 software,and the specific restriction enzyme sites were calculated.Mva Ⅰ was finally selected as a restriction endonuclease to digest the fragments of the gene.At the same time,Cyprinus carpio L.was introduced as an outgroup to construct a phylogenetic tree.It was shown from our analysis that we could distinguish the four different varieties of Carassius auratus rapidly and accurately based on polymerase chain reaction namely analysis of restriction fragment length polymorphism,which provided a theoretical basis for further study on species identification,produced a molecular method on mtDNA ND3/ND4L/ND4 to identify C.auratus auratus of Zhejiang province and C.auratus gibelio for the first time.However,only one base difference between the two among C.auratus auratus,C.auratus auratus goldfish,and C.aurtus color variety,which means their similarity close to 100%,it was failed to find any restriction endonuclease can different the three based on the analysis of the digestion software（RESearch-version 1.0）,which also verified the origin of C.auratus auratus goldfishandC.aurtus color varietyfromC.auratus auratus.

关 键 词：鲫鱼 品系 线粒体DNA PCR-RFLP 物种鉴定 

分 类 号：S961[农业科学—水产养殖]