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青海省羊源细粒棘球蚴EG95基因的克隆与序列分析

韩秀敏1，2，3， 史大中1， 贾万忠2， 杨永海3， 王虎3

(1. 兰州医学院， 甘肃 兰州730000； 2. 中国农业科学院 兰州兽医研究所， 甘肃 兰州730046；
3. 青海省疾病预防控制中心， 青海 西宁811602）

摘要： 从青海省羊源细粒棘球蚴中提取基因组DNA，用EG95特异性引物进行PCR扩增，并克隆至pGEMT Easy载体上，经PCR、EcoRⅠ酶切和测序鉴定后，用DNAstar软件对3个克隆的DNA序列进行同源性分析。结果表明，EG95基因的3个序列(EG95QH1、EG95QH2和EG95QH3)的片段大小为1435～1437bp，与GenBank中的EG95基因同源性为73.2%～99.2%，差异集中在内含子区域。其中EG95QH1、EG95QH3与GenBank EG95XJag、EG954序列的同源性较高；EG95QH2与GenBank中EG951、 EG952、 EG953序列的同源性较高。研究结果表明，青海羊源细粒棘球蚴EG95基因的3个序列为EG95基因家族的成员。

关键词： 细粒棘球绦虫； EG95基因； 克隆； 序列分析

中图分类号：S 852.734  文献标识码： A  文章编号：1000-6419(2004)01-0017-05

Cloning and analysis of EG95 gene of Echinococcus granulosus
from Qinghai Province

HAN Xiumin1,2,3, SHI Dazhong1, JIA Wanzhong2, YANG Yonghai3, WANG Hu3

(1. Lanzhou Medical College, Lanzhou 730000,China; 2. Lanzhou Veterinary Research Institute, 
Chinese Academy of Agricultural Sciences, Lanzhou 730046,China; 3. Qinghai Center of 
Disease Prevention and Control,Xining 811602,China) 

Abstract: The genomic DNA was extracted from the sheep hydatid cyst protoscolices from Qinghai Province in China. EG95 gene was amplified with a pair of specific primers using polymerase chain reaction(PCR), and cloned into pGEMT Easy plasmid. The recombinant plasmids were characterized with PCR, EcoRⅠ enzyme digestion and sequencing. The EG95QH DNA sequences were analyzed by DNAstar biosoftware. The DNA sequences of three clone fragments revealed that EG95QH sequences belong to the same EG95 gene family with the length of 1435-1437bp and that there are 73.2%-99.2% homology with EG95 gene , varies between EG95QH and other EG95 genes in GenBank. The multiple nucleotide differences are present in the noncoding gene regions. EG95QH2 is close to EG951, EG952 and EG953, and difference from EG95QH1 and EG95QH3. EG95QH1 and EG95QH3 are close to EG95XJag and EG954.

Key words: Echinococcus granulosus; EG95 gene; sequencing; genetic variation

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