CAJ | 학술논문

目的应用DNA条形码技术对陕西省鼠疫疫区啮齿动物残体进行宿主分类鉴定,分析DNA条形码技术鉴定方法的可行性.方法收集外形经形态学鉴定的7个鼠种8个不同部位62份残体样本,提取组织中DNA,经PCR、电泳、测序及结果比对等步骤,分析线粒体细胞色素C氧化酶Ⅰ亚基(COI)基因序列.将COI基因序列利用美国国家生物技术信息中心(NCBI)运行BLAST程序进行序列同源性比较;利用邻接(NJ)法构建COI基因序列的NJ系统树.结果除3份毛发外,足部、尾巴、皮毛、肌肉、肋骨、耳朵、眼球均扩增出COI基因,扩增产物片段的大小与预期片段的大小相近(700 bp),且条带单一.DNA测序获得了59份残体标本的COI基因,其中长爪沙鼠、黑线仓鼠、子午沙鼠、三趾跳鼠、小毛足鼠、褐家鼠、五趾跳鼠序列同源性比较显示,COI基因与对应宿主所对应的基因均具有高度的同源性(99.0％,98.1％,99.8％,98.9％,99.5％,99.1％,98.3％).NJ法显示,59份残体标本的COI基因构建出7个序列的NJ系统树,同鼠种不同标本聚为一类.结论 DNA条形码技术能够对鼠疫疫区啮齿动物残体进行物种分类鉴定,鉴定结果可靠.
목적 응용DNA조형마기술대합서성서역역구교치동물잔체진행숙주분류감정,분석DNA조형마기술감정방법적가행성.방법 수집외형경형태학감정적7개서충8개불동부위62빈잔체양본,제취조직중DNA,경PCR、전영、측서급결과비대등보취,분석선립체세포색소C양화매Ⅰ아기(COI)기인서렬.장COI기인서렬이용미국국가생물기술신식중심(NCBI)운행BLAST정서진행서렬동원성비교;이용린접(NJ)법구건COI기인서렬적NJ계통수.결과 제3빈모발외,족부、미파、피모、기육、륵골、이타、안구균확증출COI기인,확증산물편단적대소여예기편단적대소상근(700 bp),차조대단일.DNA측서획득료59빈잔체표본적COI기인,기중장조사서、흑선창서、자오사서、삼지도서、소모족서、갈가서、오지도서서렬동원성비교현시,COI기인여대응숙주소대응적기인균구유고도적동원성(99.0％,98.1％,99.8％,98.9％,99.5％,99.1％,98.3％).NJ법현시,59빈잔체표본적COI기인구건출7개서렬적NJ계통수,동서충불동표본취위일류.결론 DNA조형마기술능구대서역역구교치동물잔체진행물충분류감정,감정결과가고.
Objective To identify rodent residues using DNA barcoding technology in plague areas of Shaanxi and to analyze the feasibility of DNA barcoding method.Methods DNA extraction,PCR,electrophoresis and sequence determination and alignment were used to determine the cytochrome C oxidase Ⅰ (COI) gene sequence from 62 residues of 7 species identified by morphology in 8 different parts.COI gene sequence was analyzed using BLAST software of American National Center for Biotechnology Information (NCBI) for sequence homology comparison and the phylogenetic tree was constructed by using the neighbor joining(NJ)method of COI gene sequence.Results In addition to the hair,CO I genes of the feet,the tail,the fur,the muscle,the ribs,the ear,and the eye were amplified,respectively,and the size of amplified fragment was similar to the size of the product with an expected fragment(700 bp),and the fragment was a single band.COI genes of 59 residue specimen were obtained by DNA sequencing and there were high degree of homologies between CO I gene sequences of Meriones unguiculatus,Cricetulus barabensis,Meriones meridianus,Dipus sagitta,Phodopus roborovskii,Rattus norvegicus and Allactaga sibirica and their corresponding host genes in NCBI(99.0％,98.1％,99.8％,98.9％,99.5％,99.1％,98.3％).NJ method showed that 7 NJ phylogenetic trees were constructed with COI sequence of 59 species residues.The CO I sequences of same rodent with different residues were clustered into one group.Condusion DNA barcoding technology can identify host animal residues in plague areas,and the identification results are reliable.