青海省羊源细粒棘球蚴EG95基因的克隆与序列分析  

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

作　　者：韩秀敏[1] 史大中[1] 贾万忠[2] 杨永海[3] 王虎[3] 

机构地区：[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

出　　处：《中国兽医科技》2004年第1期17-21,共5页Chinese Journal of Veterinary Science and Technology

基　　金：瑞典国际基金项目 (B/ 2 85 7 1)

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 TEasy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 14 35～ 14 37bp ,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GenBankEG95XJ 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。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 1 435-1 437 bp 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.

关 键 词：青海 羊 细粒棘球蚴 EG95基因 克隆 序列分析 DNA 同源性分析 包虫病 棘球蚴病 

分 类 号：S852.734[农业科学—基础兽医学] S858.265.9[农业科学—兽医学]