Identification of gadoid species (Pisces, Gadidae) by PCR-RFLP analysis.
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.
Aranishi
Futoshi
F
Marine Molecular Ecology Laboratory, Department of Biological and Environmental Sciences, Miyazaki University, Miyazaki 889-2192, Japan. aranishi@cc.miyazaki-u.ac.jp
Okimoto
Takane
T
Izumi
Shotaro
S
D000818:Animals; D001483:Base Sequence; D004247:DNA; D048388:Gadiformes / Q000145:classification* / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012689:Sequence Homology, Nucleic Acid
D004247:DNA