基于线粒体16S rRNA基因PCR-RFLP分析的带鱼种类（鱼类：带鱼科）分子鉴定

Molecular identification of hairtail species (Pisces: Trichiuridae) based on PCR-RFLP analysis of the mitochondrial 16S rRNA gene.

作者信息

Chakraborty Anirban, Aranishi Futoshi, Iwatsuki Yukio

机构信息

United Graduate School of Agricultural Sciences, Kagoshima University, Japan.

出版信息

J Appl Genet. 2005;46(4):381-5.

PMID:16278511

Abstract

A rapid PCR-RFLP analysis was designed to identify 3 closely related species of hairtails: Trichiurus lepturus, T. japonicus, and Trichiurus sp. 2, basing on partial sequence data (600 bp) of the mitochondrial DNA encoding the 16S ribosomal RNA (16S rRNA) gene. Restriction digestion analysis of the unpurified PCR products of these 3 species, using EcoRI and VspI endonucleases, generated reproducible species-specific restriction patterns showing 2 fragments (250 bp and 350 bp) for T. lepturus in EcoRI digestion and 2 fragments (196 bp and 404 bp) for T. japonicus in VspI digestion, whereas no cleavage was observed for Trichiurus sp. 2 in both EcoRI and VspI digestions. The PCR-RFLP technique developed in this study proved to be a rapid, reliable and simple method that enables easy and accurate identification of these 3 closely related species of the genus Trichiurus.

摘要

基于编码16S核糖体RNA（16S rRNA）基因的线粒体DNA部分序列数据（600 bp），设计了一种快速聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）方法，用于鉴定3种亲缘关系密切的带鱼：小带鱼、日本带鱼和带鱼2号。使用EcoRI和VspI内切酶对这3个物种未经纯化的PCR产物进行限制性酶切分析，产生了可重复的物种特异性限制性图谱，EcoRI酶切时小带鱼显示2条片段（250 bp和350 bp），VspI酶切时日本带鱼显示2条片段（196 bp和404 bp），而带鱼2号在EcoRI和VspI酶切中均未观察到切割。本研究开发的PCR-RFLP技术被证明是一种快速、可靠且简单的方法，能够轻松准确地鉴定带鱼属的这3个亲缘关系密切的物种。