熊科动物COX2基因DNA条形码鉴定方法的建立  

Establishment of an Ursidae Identification Method through COX2 Gene Base on DNA Barcoding

作　　者：邵建宏 蒋晓霞 张洪 刘云国 麻丽丹 莫秋华 蔡一村 罗宝正 SHAO Jian-Hong;JIANG Xiao-Xia;ZHANG Hong;LIU Yun-Guo;MA Li-Dan;MO Qiu-Hua;CAI Yi-Cun;LUO Bao-Zheng(Technical Center of Gongbei Customs,Zhuhai 519001;Collegeof LifeSciences,LinyiUniversity,Linyi 276012;Shenzhen Bioeasy Biotechnology Co.,Ltd.,Shenzhen 518101;Technical Center for Animal,Plant and Food Inspection and Quarantine,Shanghai Customs,Shanghai 200135)

机构地区：[1]拱北海关技术中心,珠海519001 [2]临沂大学生命科学学院,临沂276012 [3]深圳市易瑞生物技术股份有限公司,深圳518101 [4]上海海关动植物与食品检验检疫技术中心,上海200135

出　　处：《中国口岸科学技术》2025年第2期80-88,共9页China Port Science and Technology

基　　金：海关总署科研项目(2022HK003)。

摘　　要：为有效鉴定熊科(Ursidae)物种,根据熊科动物线粒体细胞色素c氧化酶2(cytochrome c oxidase subunit 2,COX2)基因设计扩增片段为790 bp的通用引物,对9份熊科动物组织样本、4份熊科动物合成质粒和16份对照样本进行聚合酶链式反应(Polymerase Chain Reaction,PCR)扩增、电泳和测序,计算物种遗传距离,并采用邻接法构建进化树。结果显示:16份对照样本均未见扩增条带;9份熊科动物组织DNA和4份熊科动物合成质粒的检测结果符合预期,且与美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)参考序列的一致性均>96%;8种熊科动物的最大种内遗传距离为0.014%,最小种间遗传距离为0.022%,存在显著条形码差距。结果表明,本研究所建立的COX2基因DNA条形码方法能够准确鉴定熊科物种,可为海关等执法部门打击违法犯罪行为、履行《濒危野生动植物种国际贸易公约》提供技术支撑。To effectively identify the species of Ursidae,a universal primer with an amplification fragment of 790 bp was designed based on the mitochondrial cytochrome c oxidase 2(COX2)gene of Ursidae,and nine Ursidae tissue samples,four Ursidae synthetic plasmids,and sixteen control samples were subjected to polymerase chain reaction(PCR)amplification,electrophoresis,and sequencing.Genetic distances of the species were calculated,and an evolutionary tree was constructed using the neighbor-joining method.The results showed that no amplified bands were observed in the eighteen control samples.In contrast,the nine Ursidae tissue DNAs and the four Ursidae synthetic plasmids exhibited expected detection results,and the consistency with the NCBI reference sequences was>96%.Moreover,the maximum intraspecies genetic distance of the eight species of Ursidae was 0.014%,while the minimum interspecies genetic distance was 0.022%,indicating a significant barcoding gap.The study demonstrates that this DNA barcoding method for the COx2 gene can accurately identify the species of Ursidae,which can provide technical support for Customs and other law enforcement departments to combat illegal and criminal behaviors and fulfill the Convention on International Trade in Endangered Species of Wild Fauna and Flora(CITES).

关 键 词：熊 DNA条形码 细胞色素c氧化酶2 鉴定 

分 类 号：D918.9[政治法律—法学] S851.34[农业科学—预防兽医学]