利用SNP分子标记对阿根廷滑柔鱼和科氏滑柔鱼的分子鉴定

Molecular authentication of Illex argentinus and Illex coindetii by using SNP markers

目的:建立快速、准确的鉴定阿根廷滑柔鱼及其混淆品科氏滑柔鱼的DNA分子标记方法。方法:提取阿根廷滑柔鱼及科氏滑柔鱼的基因组DNA,利用PCR扩增2种鱿鱼品种的细胞色素C氧化酶亚基I(Cytochrome Oxidase Subunit I,COI)的核苷酸序列,利用序列比对发掘2种鱿鱼的品种特异性位点。根据2种鱿鱼品种的SNP位点,通过引入错配碱基设计品种特异性引物,并建立鉴定2种鱿鱼品种的多重PCR体系。结果:建立的多重PCR反应体系使阿根廷滑柔鱼扩增出了516 bp的特异性条带,科氏滑柔鱼产生了419 bp的特异性条带,2种鱿鱼的混合品则同时扩增出了516 bp和419 bp的条带。结论:利用SNP分子标记建立的多重PCR体系,快速、准确、特异性强,可作为阿根廷滑柔鱼及其加工产品原料来源鉴定的分子手段。

Objective： The aim of this study was to establish a rapid and accurate method for molecular identification of two squid species, Illex argentinus and its adulterant Illex coindetii. Methods： The nucleotide sequences of mitochondrial cytochrome oxidase subunit I（COI） were amplified by PCR from genomic DNA of two squid species. Species-specific SNP sites of two squid species were mined by multiple sequence alignments. Based on the SNP sites of two species, specific primers were designed by introducing additional mismatches, and a multiplex PCR assay was established for molecular authentication of I. argentinus and I. coindetii. Results： Multiplex PCR result showed that I. argentinus produced a 516 bp-specific band, I. coindetii yielded a 419 bp fragment, and the mixture of the two species yielded both 516 bp and 419 bp amplicons. Conclusion： A rapid and accurate DNA method was developed for molecular discrimination of I. argentinus and I. coindetii., and the method can be used for origin authentication of I. argentines and its processed products.