Polymorphism of cox1 and nad1 of Echinococcus granulosus in free-range yaks and sheep in central Xizang
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 ampli-fied 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 ana-lyzed 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 bas-es in the cox1 sequence was 32.6%,33.1%,17.4%and 16.9%respectively.The average content of A,T,G and C bas-es in the nad1 sequence was 19.5%,47.6%,25.5%and 7.4%.The results of homology analysis showed that the ho-mology between cox1 sequences was 99.6%to 100.0%,and the homology between nad1 sequences was 98.1%to 100.0%.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 granu-losus 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 granulo-sus in central Xizang.
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