布氏田鼠疫源地鼠疫宿主动物DNA条形码分析  

Identification of host animals for plague by DNA barcoding in the epidemic focus of Lasiopodomys brandti

作　　者：常子丽[1] 张忠兵[1] 刘芳[1] 王建军[1] 马志东[1] 李振林[1] 赵成祥[1] 杨晓娟[1] 范蒙光[1] 

机构地区：[1]内蒙古自治区地方病防治研究中心中心实验室,呼和浩特010031

出　　处：《中华地方病学杂志》2014年第6期605-608,共4页Chinese Journal of Endemiology

摘　　要：目的 探究应用DNA条形码技术对鼠疫宿主动物进行分子鉴定的可行性.方法 采集内蒙古自治区锡林郭勒高原布氏田鼠鼠疫自然疫源地内的7种啮齿动物,包括布氏田鼠、达乌尔黄鼠、达乌尔鼠兔、蒙古毛足鼠、五趾跳鼠、长爪沙鼠和大沙鼠,提取肝脏组织,进行线粒体细胞色素氧化酶Ⅰ （CO Ⅰ）基因序列分析,用MEGA 5.0软件计算GC含量,通过双参数（Kimura-2-parameter,K2P）模型计算遗传距离,采用邻接法（Neighbor-Joining,NJ）构建系统发育树.结果 共获得61条CO Ⅰ基因序列片段,每条序列长度657 bp.五趾跳鼠、布氏田鼠、长爪沙鼠、达乌尔鼠兔、蒙古毛足鼠、大沙鼠、达乌尔黄鼠的GC含量分别为（46.945±0.109）％、（43.836±0.000）％、（40.313±0.084）％、（47.514±0.073）％、（41.838±0.073）％、（40.335±0.000）％、（39.467±0.032）％,除大沙鼠和长爪沙鼠的GC含量组间比较差异无统计学意义（P＞ 0.05）,其他各鼠种的GC含量组间比较差异均有统计学意义（P均＜ 0.05）.上述7个鼠种在第1位密码子处GC含量分别为（57.534±0.000）％、（55.251±0.000）％、（54.338±0.000）％、（57.078±0.000）％、（55.308±0.057）％、（53.881±0.000）％、（56.164±0.000）％,除布氏田鼠和蒙古毛足鼠的GC含量组间比较差异无统计学意义（P＞ 0.05）,其他各鼠种的GC含量组间比较差异均有统计学意义（P均＜0.05）;上述7个鼠种在第3位密码子处GC含量分别为（40.835±0.328）％、（33.790±0.000）％、（24.136±0.253）％、（43.455±0.218）％、（27.740±0.207）％、（24.201±0.000）％、（19.772±0.097）％,除大沙鼠和长爪沙鼠的GC含量组间比较差异无统计学意义（P＞0.05）,其他各鼠种的GC含量组间比较差异均有统计学意义（P均＜0.05）.各鼠种的种内平均遗传距离为0.003,种间平均遗传距离为0.234 8,NJ系统发育树能清楚的区分7个不同的类群Objective To study the feasibility of DNA barcoding in identification of host animals with plague.Methods Seven species were collected,including Lasiopodomys brandti,Spermophilus dauricus,Ochotona daurica,Phodopus campbelli,Allactaga sibirica,Meriones unguiculatus and Rhombomys opimus in the epidemic focus of Lasiopodomys brandti,Inner Mongolia,China.Liver was extracted and mitochondrial cytochrome oxidase subunit Ⅰ gene （CO Ⅰ） was analyzed.The GC content and genetic distance by Kimura-2-parameter（K2P） were calculated using MEGA5.0,and the phylogenetic trees were constructed with Neighbor-Joining （NJ） method.Results Sixty-one CO Ⅰ gene sequence fragments were barcoded,and each sequence was 657 bp in length.The GC% of A llactaga sibirica,Lasiopodomys brandti,Meriones unguiculatus,Ochotona daurica,Phodopus campbelli,Rhombomys opimus and Spermophilus dauricus were （46.945 ± 0.109）%,（43.836 ± 0.000）%,（40.313 ± 0.084）%,（47.514 ± 0.073）%,（41.838 ± 0.073）%,（40.335 ± 0.000）% and （39.467 ± 0.032）%,respectively,and the differences between them were significant（all P 〈 0.05）,except the difference between Rhombomys opimus and Meriones unguicalatus（P 〉 0.05）.The 1st codon GC contents of the seven species were （57.534 ± 0.000）%,（55.251 ± 0.000）%,（54.338 ± 0.000）%,（57.078 ± 0.000）%,（55.308 ± 0.057）%,（53.881 ± 0.000）% and （56.164 ± 0.000）%,respectively,and the differences between them were significant （all P 〈 0.05）,except the difference between Lasiopodomys brandti and Phodopus campbelli （P 〉 0.05）; the 3rd codon GC content of the seven species were （40.835 ± 0.328）%,（33.790 ± 0.000）%,（24.136 ± 0.253）%,（43.455± 0.218）%,（27.740 ± 0.207）%,（24.201 ± 0.000）% and （19.772 ± 0.097）%,respectively,and the differences were significant （all P 〈 0.05） among species,except the difference between Rhombomys opimus and Meriones unguicalatus （P 〉 0.05）.The average intraspecific genetic di

关 键 词：DNA条形码 COⅠ基因 鼠疫 

分 类 号：R516.8[医药卫生—内科学]