利用PCR和限制性酶切技术鉴别3种鳗鱼-《集美大学学报（自然科学版）》

文章信息/Info

Title:: Identification of Three Eel Species with PCR and Restriction Analysis

作者:: 刘光明1; 2; 史千玉1; 2; 曹敏杰1; 2; 苏国成1; 2; 倪辉1; 2; 蔡慧农1; 2; (1.集美大学生物工程学院,福建厦门 361021;2.福建省高校水产科学技术与食品安全重点实验室,福建厦门 361021)

Author(s):: LIU Guang-ming1; 2; SHI Qian-yu1; 2; CAO Min-jie1; 2; SU Guo-cheng1; 2; NI Hui1; 2; CAI Hui-nong1; 2; (1.School of Biotechnology Engineering,Jimei University,Xiamen 361021,China2.Key Laboratory of Science andTechnology for Aquaculture and Food Safety(Jimei University),Fujian Province,Xiamen 361021China)

Keywords:: eel; 16S rRNA; PCR; restriction analysis; molecular identification

摘要:: 根据日本鳗、欧洲鳗和美洲鳗3种鳗鱼的16S rRNA基因序列，设计特异性引物，进行16S rRNA基因的PCR扩增，然后对PCR产物进行Apa I酶切反应.结果显示，日本鳗和欧洲鳗的PCR产物均出现211 bp的特异性扩增条带，美洲鳗的PCR产物出现两条DNA条带；日本鳗的PCR产物经Apa I酶切产生大小为135 bp和76 bp的两条条带，而欧洲鳗的PCR产物经Apa I酶切没有变化.因此，利用该方法可鉴别出日本鳗、欧洲鳗和美洲鳗等3种鳗鱼

Abstract:: According to the sequences of 16S rRNA genes of three eels(Anguilla japonica, Anguilla anguilla,and Anguilla rostrata),a pair of primers were designed for amplification of partial 16S rRNA genes. The PCR products were recycled and digested with restriction enzyme Apa I.Electrophoretic analysis of the PCR products from A.japonica and A.anguilla did not indicate any difference of one band in length of 211 bp,but the PCR products from A.rostrata had two bands.Since only partial 16S rRNA gene of A. japonica had the restriction site of Apa I,two fragments of 135 bp and 76 bp were presented only for the PCR products from A.japonica through Apa I,while the PCR products from A.anguilla were not digested with Apa I.PCR amplification and Apa I restriction analysis of partial 16S rRNA gene of A.japonica, A.anguilla and A.rostrata were suitable genetic markers to distinguish three eel species.