通过聚合酶链反应-限制性片段长度多态性分析鉴定鳕科鱼类（硬骨鱼纲，鳕科）

Identification of gadoid species (Pisces, Gadidae) by PCR-RFLP analysis.

作者信息

Aranishi Futoshi, Okimoto Takane, Izumi Shotaro

机构信息

Marine Molecular Ecology Laboratory, Department of Biological and Environmental Sciences, Miyazaki University, Miyazaki 889-2192, Japan.

出版信息

J Appl Genet. 2005;46(1):69-73.

PMID:15741666

Abstract

A rapid PCR-RFLP analysis was optimized to identify the presence of 3 closely related gadoid fish species: Alaska pollack Theragra chalcogramma, Pacific cod Gadus macrocephalus and Atlantic cod Gadus morhua in commercial seafood products. Gadoid universal primers were designed for PCR amplification of a 558-bp fragment encoding the mitochondrial cytochrome b gene. Without purification of the PCR products, double digestion with Eco32I and Eco105I restriction enzymes generated reproducible species-specific restriction patterns visualizing 3 fragments (106 bp, 161 bp and 291 bp) in Alaska pollack and 2 fragments (106 bp and 452 bp) in Pacific cod, whereas no cleavage was observed in Atlantic cod. This PCR-RFLP analysis is simple, rapid and reliable, and therefore can be routinely applied to discover fraudulent substitution among 3 economically important gadoid species in commercial seafood products.

摘要

优化了一种快速聚合酶链反应-限制性片段长度多态性分析方法，以鉴定商业海鲜产品中3种亲缘关系密切的鳕科鱼类的存在：狭鳕鱼（Theragra chalcogramma）、太平洋真鳕（Gadus macrocephalus）和大西洋鳕鱼（Gadus morhua）。设计了鳕科通用引物，用于聚合酶链反应扩增编码线粒体细胞色素b基因的558碱基对片段。无需纯化聚合酶链反应产物，用Eco32I和Eco105I限制性内切酶进行双酶切，可产生可重复的物种特异性限制性图谱，狭鳕鱼显示3个片段（106碱基对、161碱基对和291碱基对），太平洋真鳕显示2个片段（106碱基对和452碱基对），而大西洋鳕鱼未观察到切割。这种聚合酶链反应-限制性片段长度多态性分析方法简单、快速且可靠，因此可常规用于发现商业海鲜产品中3种经济上重要的鳕科鱼类之间的欺诈性替代情况。