摘要
由烟台海鲜市场随机采集具有相对重要商业价值的10种海鱼(小黄鱼、牙片、鲅鱼、面条鱼、鲐鱼、鲳鱼、鲈鱼、偏口、黑鱼、白姑鱼),利用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)技术对鱼种进行鉴定.首先提取10种海鱼的基因组DNA,利用自行设计的通用引物对其线粒体细胞色素C氧化酶亚基Ⅲ基因(MT-co3)进行PCR扩增,对扩增产物克隆、测序,用NCBI数据库检索,确定了10种海鱼的鱼种.依据测序结果进行限制性内切酶酶切分析,选用NlaⅢ和HinfⅠ2种内切酶进行酶切,获得了各种鱼类特异的酶切片段图谱.研究结果表明,PCR-RFLP能有效地区分以上10种海鱼,在鱼种鉴定上具有简便、准确的优势,可为鱼类食品检测提供科学依据.
The PCR-based restriction fragment length polymorphism（ PCR-RFLP） is used to identify 10 marine fish species（ Larimichthys polyactis,Paralichthys olivaceus,Scomberomorus niphonius,Ammodytes personatus,Scomber japonicus,Peprilus medius,Lateolabrax japonicus,Cleisthenes herzensteini,Sebates schlegeli,Pennahia argentata,） with relatively important commercial value,which are randomly sampled from Yantai seafood market. The genomic DNAs of the 10 species of marine fish are extracted,and the mitochondria cytochrome c oxidase subunit Ⅲ gene（ MT-co3） of all samples are amplified with a pair of self-designed universal primers. The PCR products are then cloned and sequenced. RFLP analysis is carried out based on the sequencing result using two restriction enzymes,NlaⅢ and HinfⅠ,to digest the PCR products. A specific-restriction map for each fish species is obtained through the PCR-RFLP method. The results show that the PCR-RFLP is a simple and accurate method that can effectively distinguish the 10 selected fish species. This method may provide scientific basis for fish food source testing.
出处
《烟台大学学报（自然科学与工程版）》
CAS
2015年第2期97-102,共6页
Journal of Yantai University(Natural Science and Engineering Edition)
基金
山东省自然科学基金资助项目(ZR2013CQ021)
关键词
聚合梅链式反应-限制性片段长度多态性技术
物种鉴定
线粒体细胞色素C氧化酶亚基Ⅲ基因
海鱼
polymerase chain reaction-based restriction fragment length polymorphism（PCR-RFLP）
species identification
cytochrome c oxidase subunit Ⅲ gene of mitochondrion DNA（MT-co3）
marine fish
