应用PCR-RFLP和芯片生物分析系统鉴别河豚鱼品种

摘要

应用聚合酶链式反应-限制性片段长度多态分析和芯片生物分析系统建立5种河豚的分子生物学品种鉴定新方法。首先提取鱼的DNA进行细胞色素b基因的PCR扩增，然后用DdeⅠ、Hae Ⅲ和Nla Ⅲ三种限制性内切酶进行酶切，在Agilent DNA1000芯片上对酶切片段进行分离。该方法可成功区分5个河豚鱼品种，是一种快速、简便、有效的鱼类品种鉴定分析手段。%Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and chip bio-analysis system were used to identify ifve puffer species. A 464-bp fragment of mitochondrial cytochrome b gene was ampliifed by PCR and the products were digested with the restriction endonuclease DdeⅠ, HaeⅢand NlaⅢ, respectively. The resulting fragments were further resolved on DNA chip. Five puffer species were successfully identiifed by this method. In conclusion, PCR-RFLP and chip bio-analysis system could provide a fast, easy, automatic and reliable analysis approach.