牦牛病毒性腹泻病毒E1基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of the E1 Gene of Bovine Viral Diarrhea Virus Strain Yak

作　　者：王文伯[1] 刘亚刚[1] 胡炳峰[1] 杨晓农[1] 于吉锋[2] 冯英阳[1] 王盼盼[1] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041 [2]四川省畜牧科学研究院,成都610041

出　　处：《中国畜牧兽医》2010年第3期94-98,共5页China Animal Husbandry & Veterinary Medicine

基　　金：四川省科技厅攻关项目(04ZY029-006-04);国家民委项目(07XN03)

摘　　要：参考GenBank中发表的BVDV毒株的基因组序列设计2对引物,利用套式RT-PCR方法首次成功克隆牦牛体内分离鉴定出的牛病毒性腹泻病毒E1基因,并扩增出预期的585 bp目的片段。将扩增产物克隆至pMD18-T Vector,经质粒PCR鉴定及酶切鉴定获得阳性重组质粒并进行测序。测序结果经BLAST同源性比较分析,克隆得到的E1基因与Osloss株同源性最高,但核苷酸同源性仅为73.3%,推导氨基酸同源性仅为82.6%,表明牦牛病毒性腹泻病毒存在较大的基因突变。这可能是该病毒为适应牦牛这种特有生物体和牦牛所生活的高原生态环境的结果,或该病毒也可能具有独立的遗传衍化来源。According to the sequence data of BVDV strain published by GenBank,two set of primers were designed and used to amplify E1 gene of BVDV strain yak by the method of nested reverse transcription polymerase chain reaction（RT-PCR）.A specific 585 bp DNA segment was amplified,which was cloned into pMD18-T vector.The positive recombinant clone was identified by plasmid PCR and restriction enzyme digestion.The recombinant plasmid was sequenced and compared with BLAST on line.The E1 gene of BVDV strain yak exhibits the highest homology with strain Osloss,but they only shares 73.3% nucleotide sequence identity and 82.6% amino acid identity,showing that the E1 gene of BVDV strain yak have great variation.This may be the virus to adapt to yak and the ecological environment of the plateau,or the virus may also have an independent source of genetic.

关 键 词：牛病毒性腹泻病毒 牦牛 E1基因 克隆 生物信息学分析 

分 类 号：Q78[生物学—分子生物学]