›› 2010, Vol. 37 ›› Issue (3): 94-98.

• 生物技术 • 上一篇    下一篇

牦牛病毒性腹泻病毒E 1基因的克隆及生物信息学分析

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   

  1. (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)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-03-20 发布日期:2010-03-20
  • 通讯作者: 刘亚刚

Cloning and Bioinformatics Analysis of the E 1 Gene of Bovine Viral Diarrhea Virus Strain Yak

WANG Wen-bo1, LIU Ya-gang1, HU Bing-feng1, YANG Xiao-nong1, 
YU Ji-feng2, FENG Ying-yang1, WANG Pan-pan1
  

  1. (1.College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China; 2.Sichuan Animal Science Academy, Chengdu 610041, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-03-20 Published:2010-03-20
  • Contact: LIU Ya-gang

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E 1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E 1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。

关键词: 牛病毒性腹泻病毒; 牦牛; E 1基因; 克隆; 生物信息学分析

Abstract: According to the sequence data of BVDV strain published by GenBank, two set of primers were designed and used to amplify E 1 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 E 1 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 E 1 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.

Key words: bovine viral diarrhea virus; Yak; E 1 gene; cloning; bioinformatics analysis

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