基于COI基因的赣北地区小泡巨鼠的鉴定  

Identification of Leopoldamys edwardsi in Northern Jiangxi based on COI gene

作　　者：陈飞 胡海军 刘占斌 陶卉英 钱科 郑卫青 CHEN Fei;HU Haijun;LIU Zhanbin;TAO Huiying;ZHENG Weiqing(National Key Laboratory for the Prevention and control of infectious diseases,Key Laboratory of Animal Origin and Vector-borne Infectious Diseases of Jiangxi Province,Department for Disinfection and Vector Control,Nanchang Center for Disease Control and Prevention,Nanchang 330038,Jiangxi,China;Hainan Medical College,School of Tropical Medicine,Haikou 571199;Nanchang Police Dog Base of the Ministry of Public Security)

机构地区：[1]传染病预防控制国家重点实验室研究基地,江西省动物源与媒介生物性传染病重点实验室,南昌市疾病预防控制中心消毒与病媒生物防制科,江西南昌330038 [2]海南医学院,热带医学院,海南海口571199 [3]公安部南昌警犬基地,江西南昌330100

出　　处：《中华卫生杀虫药械》2023年第6期495-498,共4页Chinese Journal of Hygienic Insecticides and Equipments

基　　金：南昌市医疗技术支撑计划重点项目(2022-KJZC-024);南昌市医疗技术支撑计划一般项目(2022-KJZC-018)

摘　　要：目的以线粒体细胞色素C氧化酶亚基(COI)基因为目的基因,利用DNA条形码技术对未知鼠样本进行鉴定.方法采用夹夜法对九江市武宁县宋溪镇某村的鼠密度进行调查,经形态学初步鉴定后,以捕获的1只无法明确鉴定的大型鼠样本为对象,提取基因组DNA并采用COI基因通用引物BatL5310和R6036R进行PCR扩增及测序,将测序结果在NCBI网站进行BLAST比对,采用MEGA7软件选择最大似然法构建系统发育树.结果共捕获小型兽类7只,密度为3.38%,经形态学鉴定有褐家鼠2只,黑线姬鼠2只,臭鼩鼱2只,形态学初步鉴定为青毛巨鼠或小泡巨鼠的未知鼠1只,提取的未知鼠样本DNA通过PCR成功扩增出COI基因目的条带,与NCBI基因库中小泡巨鼠(KP995219)序列同源性高达100%,系统发育树显示该鼠样本与小泡巨鼠(KP995219、KP995203)处于同一分支,遗传距离分别为0、0.0016,而青毛巨鼠(NC_036730)单独处于另一分支,遗传距离较远.结论赣北地区首次报道小泡巨鼠,采用COI基因的DNA条形码技术可快速、准确的进行鼠种鉴定,有助于弥补非常见鼠形态学鉴定困难的不足.Objective To identify unknown rodentsample by DNA barcoding technology with mitochondrial cytochrome C oxidase subunit(COI)gene as target gene.Methods The density of rodents in a village of Songxi Town,Wuning County,Jiujiang City was determined by the trapping method.Following preliminary morphological identification,a rare large rodent specimen was captured and selected for genomic DNA extraction and the COI gene was amplified using universal primers(BatL5310 and R6036R),then sequenced.BLAST comparison was subsequently carried out in NCBI.The phylogenetic tree was calculated by the Maximum Likelihood method using MEGA7software.Results A total of seven small mammals were captured in northern Jiangxi and the density was 3.38%.Morphological identification revealed the presence of 2 Rattusnorvegicu,2 Apodemus agrarianus,and 2 Suncus murinus.Additionally,a preliminary morphological identification classified one specimen as an unknown rodent,potentially belonging to the species of Berylmys bowersi or Leopoldamys edwardsi.The DNA extracted from the unknown rodent sample successfully amplified the target band of the COI gene through PCR.The COI gene showed high sequence similarities(100%)with the sequences of Leopoldamys edwardsi(KP995219)depoisted in GenBank Sequences of unknown rodent sample and Leopoldamys edwardsi(KP995219,KP995203)were clusteredin the same branch at the phylogenetic tree,and genetic distance was 0,0.0016 respectively.In addition,the Berylmys bowersi(NC_036730)was in another branch alone,and genetic distance was big.Conclusion Leopoldamys edwardsi was initially captured in northern Jiangxi.DNA barcoding is a highly effective approach to rapidly identify rodent species,and useful for the molecular identification of uncommon rodent species.

关 键 词：小泡巨鼠 DNA条形码 细胞色素C氧化酶亚基Ⅰ基因 系统发育 

分 类 号：R184.3[医药卫生—流行病学]