이중-PCR을 이용한 홍합, Mytilus coruscus의 신속한 종 판별법 개발

Abstract

Using multiplex polymerase chain reaction (PCR) with species-specific primers, a rapid method was developed to distinguish between the hard-shelled mussel (Mytilus coruscus) and Mediterranean mussel (M. galloprovincialis) in order to prevent their illegal distribution, and they are morphologically similar to each other during the spat period. The designed primers amplify the mitochondrial cytochrome b (COB) region from mussel shellfish (genus Mytilus). Using the sequences of the COB region, the Mytilus_c primer set that shows amplification for mussel shellfish (genus Mytilus) and the Mytilus_corS primer set that shows specific amplification for hard-shelled mussel was developed. In the hard-shelled mussel, duplex-PCR with both primer sets amplified a species-specific band of 334 bp with a 236 bp band common to the mussel genus. However, in the Mediterranean mussel, only a common 236 bp band was amplified. This result suggests that duplex-PCR with two primer sets allows identification of errors that may occur in the PCR process (i.e., lack of bands) and clearly distinguishes between hard-shelled mussel and Mediterranean mussel using a single PCR. Therefore, the molecular markers developed in this study can be a powerful and accurate tool to rapidly distinguish between hard-shelled mussels and Mediterranean mussel.