中华地方病学杂志
中華地方病學雜誌
중화지방병학잡지
Chinese Journal of Endemiology
2014年
6期
605-608
,共4页
常子丽%张忠兵%刘芳%王建军%马志东%李振林%赵成祥%杨晓娟%范蒙光
常子麗%張忠兵%劉芳%王建軍%馬誌東%李振林%趙成祥%楊曉娟%範矇光
상자려%장충병%류방%왕건군%마지동%리진림%조성상%양효연%범몽광
DNA条形码%COⅠ基因%鼠疫
DNA條形碼%COⅠ基因%鼠疫
DNA조형마%COⅠ기인%서역
DNA barcoding%CO Ⅰ gene%Plague
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.结论 应用DNA条码技术检测COⅠ基因,可以对内蒙古布氏田鼠疫源地的鼠疫宿主动物进行鉴别,同时可为形态鉴别提供佐证数据.
目的 探究應用DNA條形碼技術對鼠疫宿主動物進行分子鑒定的可行性.方法 採集內矇古自治區錫林郭勒高原佈氏田鼠鼠疫自然疫源地內的7種齧齒動物,包括佈氏田鼠、達烏爾黃鼠、達烏爾鼠兔、矇古毛足鼠、五趾跳鼠、長爪沙鼠和大沙鼠,提取肝髒組織,進行線粒體細胞色素氧化酶Ⅰ (CO Ⅰ)基因序列分析,用MEGA 5.0軟件計算GC含量,通過雙參數(Kimura-2-parameter,K2P)模型計算遺傳距離,採用鄰接法(Neighbor-Joining,NJ)構建繫統髮育樹.結果 共穫得61條CO Ⅰ基因序列片段,每條序列長度657 bp.五趾跳鼠、佈氏田鼠、長爪沙鼠、達烏爾鼠兔、矇古毛足鼠、大沙鼠、達烏爾黃鼠的GC含量分彆為(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)％,除大沙鼠和長爪沙鼠的GC含量組間比較差異無統計學意義(P＞ 0.05),其他各鼠種的GC含量組間比較差異均有統計學意義(P均＜ 0.05).上述7箇鼠種在第1位密碼子處GC含量分彆為(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)％,除佈氏田鼠和矇古毛足鼠的GC含量組間比較差異無統計學意義(P＞ 0.05),其他各鼠種的GC含量組間比較差異均有統計學意義(P均＜0.05);上述7箇鼠種在第3位密碼子處GC含量分彆為(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)％,除大沙鼠和長爪沙鼠的GC含量組間比較差異無統計學意義(P＞0.05),其他各鼠種的GC含量組間比較差異均有統計學意義(P均＜0.05).各鼠種的種內平均遺傳距離為0.003,種間平均遺傳距離為0.234 8,NJ繫統髮育樹能清楚的區分7箇不同的類群.結論 應用DNA條碼技術檢測COⅠ基因,可以對內矇古佈氏田鼠疫源地的鼠疫宿主動物進行鑒彆,同時可為形態鑒彆提供佐證數據.
목적 탐구응용DNA조형마기술대서역숙주동물진행분자감정적가행성.방법 채집내몽고자치구석림곽륵고원포씨전서서역자연역원지내적7충교치동물,포괄포씨전서、체오이황서、체오이서토、몽고모족서、오지도서、장조사서화대사서,제취간장조직,진행선립체세포색소양화매Ⅰ (CO Ⅰ)기인서렬분석,용MEGA 5.0연건계산GC함량,통과쌍삼수(Kimura-2-parameter,K2P)모형계산유전거리,채용린접법(Neighbor-Joining,NJ)구건계통발육수.결과 공획득61조CO Ⅰ기인서렬편단,매조서렬장도657 bp.오지도서、포씨전서、장조사서、체오이서토、몽고모족서、대사서、체오이황서적GC함량분별위(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)％,제대사서화장조사서적GC함량조간비교차이무통계학의의(P＞ 0.05),기타각서충적GC함량조간비교차이균유통계학의의(P균＜ 0.05).상술7개서충재제1위밀마자처GC함량분별위(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)％,제포씨전서화몽고모족서적GC함량조간비교차이무통계학의의(P＞ 0.05),기타각서충적GC함량조간비교차이균유통계학의의(P균＜0.05);상술7개서충재제3위밀마자처GC함량분별위(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)％,제대사서화장조사서적GC함량조간비교차이무통계학의의(P＞0.05),기타각서충적GC함량조간비교차이균유통계학의의(P균＜0.05).각서충적충내평균유전거리위0.003,충간평균유전거리위0.234 8,NJ계통발육수능청초적구분7개불동적류군.결론 응용DNA조마기술검측COⅠ기인,가이대내몽고포씨전서역원지적서역숙주동물진행감별,동시가위형태감별제공좌증수거.
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 distance (K2P) was 0.003 and the average interspecific genetic distance was 0.234 8.A NJ tree showed 7 major clusters.Conclusion CO Ⅰ sequencing of DNA barcoding can be used to identify host animals with plague in the epidemic focus of Losiopodomys brandti,and can provide stronger evidences in identification of species based on morphology.
