牦牛病毒性黏膜病病毒结构蛋白核苷酸的序列测定与分析  

Cloning and bioinformatics analysis of the structure protein gene of bovine viral diarrhea virus strain yak

作　　者：刘亚刚[1] 孙凯[1] 王妍[1] 王盼盼[1] 王荣[1] 胡炳峰[1] 王文伯[1] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041

出　　处：《西南民族大学学报（自然科学版）》2012年第4期564-569,共6页Journal of Southwest Minzu University(Natural Science Edition)

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

摘　　要：对首次从牦牛体内分离出的BVDV进行结构蛋白的研究与分析具有重要的理论价值和学术意义,本研究参考GenBank中发表的BVDV毒株的基因组序列设计四对对引物,利用PCR扩增出预期的2700bp目的片段.扩增产物克隆至pMD19-T Vector,经质粒PCR鉴定及酶切鉴定获得阳性重组质粒并对其进行测序.测序结果经DNNMAN同源性比较分析,克隆得到的结构蛋白基因与NADL株同源性最高,但核苷酸同源性仅为70.1%,推导氨基酸同源性仅为74.4%,证明牦牛株BVDV结构蛋白基因存在较大的变异.生物信息学分析表明结构序列中有片段高度疏水,蛋白基因推导氨基酸序列亲水性与抗原性成平行相关,亲水性和抗原性与标准毒株很相似.The structure protein research and analysis of the BVDV separated from the yak body for the first time is of great theoretical value and academic significance. According to the sequence data of BVDV strain published by GenBank, four sets of primers are designed and used to amplify structure protein gene of BVDV strain yak by the method of PCR. A specific 2700bp DNA segment is amplified, which is cloned into pMD19-T Vector. The positive recombinant clone is identified by plasmid PCR and restriction enzyme digestion. The recombinant plasmid is sequenced. Compared with DNAMAN, the structure protein gene of BVDV strain yak exhibits the highest homology with strain NADL, but they only share 70.1% nucleotide sequence identity and 74.4 % amino acid identity, showing that the structure protein gene of BVDV strain yak has great variation. Bioinformatics analysis shows that the structure of sequence fragments of high water, protein gene amino acid sequence with nucleic is hydrophilic into parallel related, hydrophilic and nucleic standard strain very similar.

关 键 词：牦牛病毒性腹泻病毒:结构蛋白基因 克隆测序 生物信息学分析 

分 类 号：S858.23[农业科学—临床兽医学]