西藏中部地区散养牦牛和绵羊细粒棘球绦虫cox1和nad1的多态性  

Polymorphism of cox1 and nad1 of Echinococcus granulosus in free-range yaks and sheep in central Xizang

作　　者：陈欣怡 梁宏[1] 赤列旺姆 扎西旺杰 德吉玉珍 唐文强[5] 赵海龙[1] CHEN Xinyi;LIANG Hong;Chiliewangmu;Zhaxiwangjie;Dejiyuzhen;TANG Wenqiang;ZHAO Hailong(Medical Department of Qinghai University,Department of Basic Medicine,Xining 810016,China;Nagqu Agriculture and Animal Husbandry(Grass Industry)Science and Technology Research and PromotionCenter,Nagqu 852000,China;Bango County Agriculture and Animal Husbandry Science and TechnologyService Station,Nagqu 852000,China;Dulong Deqing District Bureau of Agriculture and Rural Affairs,Lhasa 850000,China;Institute of Animal Husbandry and Veterinary Medicine,Academy of Agriculture and Animal Husbandry,Lhasa 850000,China)

机构地区：[1]青海大学医学部基础医学部,西宁810016 [2]那曲市农牧业(草业)科技研究推广中心,那曲852000 [3]班戈县农牧科学技术服务站,那曲852000 [4]堆龙德庆区农业农村局,拉萨850000 [5]西藏自治区农牧科学院畜牧兽医研究所,拉萨850000

出　　处：《中国高原医学与生物学杂志》2024年第2期107-114,共8页Journal of Chinese High Altitude Medicine & Biology

基　　金：西藏自治区科技厅重大科技专项(NO.XZ202101ZD002N)。

摘　　要：目的分析西藏中部地区散养牦牛和绵羊细粒棘球绦虫cox1、nad1的基因多态性。方法采集西藏中部地区45份散养牦牛和绵羊细粒棘球蚴包囊,提取包囊生发层DNA,根据细粒棘球蚴线粒体cox1、nad1基因序列设计扩增的引物序列,用PCR技术扩增线粒体cox1、nad1基因片段,将扩增产物做琼脂糖凝胶电泳并测序;利用MEGA11.0软件做cox1、nad1序列特征与同源性比对,并构建系统进化树;运用DnaSP 5.0软件计算序列单倍型数量(H),分析核苷酸多样性(Pi)、单倍体型多样性(Hd)特性;利用Network软件为cox1、nad1基因的所有单倍体构建单倍体网络图,判断西藏中部地区散养牦牛和绵羊的主要单倍型。结果(1)cox1、nad1序列长度分别为925、839bp。(2)在cox1序列中,A、T、G、C碱基的平均含量分别为32.6%、33.1%、17.4%、16.9%;而nad1序列中,A、T、G、C碱基的平均含量为19.5%、47.6%、25.5%、7.4%;同源性分析结果显示,cox1序列之间的同源性为99.6%~100.0%,nad1序列之间的同源性为98.1%~100.0%;西藏中部地区散养牦牛和绵羊细粒棘球绦虫基因型为G1、G3、G6。(3)cox1、nad1单倍型个数分别为19个、7个,Hd、Pi分别为0.89600、0.56000和0.36500、0.00060。(4)单倍型网络图显示,Hc-2、Hc-12、Hn-1为西藏中部地区散养牦牛和绵羊的主要单倍型。结论西藏中部地区散养牦牛和绵羊细粒棘球绦虫基因型为G1、G3、G6,G1是主要基因型。上述结论为探明西藏中部地区细粒棘球蚴的分型和病原体遗传演化的情况提供了参考依据。Objective To analyze the gene polymorphisms of cox1 and nad1 of Echinococcus granulosus in free-range yaks and sheep in central Xizang.Method 45 samples of free-range yaks and sheep hydatid cyst were collected from central Xizang.The DNA from the germinal layer of the cyst was extracted.The primers were amplified according to mitochondrial cox1 and nad1 sequences.The gene fragments of cox1 and nad1 were amplified by PCR,and the amplified products were sequenced by agarose gel electrophoresis.The software MEGA11.0 was used to compare and analyze the sequence characteristics and homology of cox1 and nad1,and construct a phylogenetic tree.The characteristics of haplotype number(H),nucleotide diversity(Pi)and haplotype diversity(Hd)were analyzed by the software DnaSP 5.0.Network software was adopted to construct haplotype network of all haplotypes of cox1 and nad1 to determine the main haplotypes of free-range yaks and sheep in central Xizang. Results(1)The sequence lengths of cox1 and nad1 were 925 and 839 bp respectively.(2)The average content of A,T,G and C bases in the cox1 sequence was 32.6%,33.1%,17.4% and 16.9% respectively.The average content of A,T,G and C bases in the nad1 sequence was 19.5%,47.6%,25.5% and 7.4%.The results of homology analysis showed that the homology between cox1 sequences was 99.6% to 100.0%, and the homology between nad1 sequences was 98.1% to100.0%.The genotypes of Echinococcus granulosus in free-range yaks and sheep in central Xizang were G1,G3 and G6.(3)The number of haplotypes of cox1 and nad1 was 19 and 7 respectively, while Hd and Pi were 0.89600,0.56000 and 0.36500, 0.00060 respectively.(4)Haplotype network indicated the main haplotypes of free-range yaks and sheep were Hc-2,Hc-12 and Hn-1 in central Xizang. Conclusion The genotypes of Echinococcus granulosus in free-range yaks and sheep in central Xizang are G1,G3 and G6,of which G1 is the main genotype,which provides reference data for exploring the classification and pathogenic genetic evolution of Echinococcus granulosus in c

关 键 词：西藏 散养 牦牛 绵羊 细粒棘球绦虫 cox1 nad1 基因 多态性 

分 类 号：S383.3[农业科学—农艺学]