水貂阿留申病毒5个分离株的全基因测序及遗传变异分析

Genome Sequencing and Genetic Variation Analysis of 5 Isolated Strains of Aleutian Mink Disease Viruses

流行病学调查表明,水貂阿留申病是严重危害水貂养殖业的3大疫病之首。迄今为止,尚未开发出能有效防控水貂阿留申病的疫苗。本试验根据GenBank上公布的不同水貂阿留申病毒(ADV)毒株的基因序列,设计并合成了12对引物,对吉林农业大学经济动物疾病实验室已分离和鉴定且具有致病性的5株水貂阿留申病毒野毒株进行全基因测序,并将测序结果与ADV参考毒株及细小病毒亚科各属代表种的全基因序列进行核苷酸序列分析及同源性比较。结果表明,所分离5株病毒基因组大小分别为4 537、4 539、4 562、4 572、4 569bp。同源性分析结果显示,分离的5株ADV毒株与欧美毒株ADV-G、ADVUtahl、ADV-SL3的核苷酸同源性最低为92.8%,最高为97.6%。通过DNA Star软件对5株ADV的VP2核苷酸推导的氨基酸序列分析可知,所分离的5个毒株中,有61个氨基酸发生突变。ADV-DL124和ADV-DL125与ADV-G株相同,在26-35位处缺失9个氨基酸,同时在36位缺失1个氨基酸(甘氨酸)。对这些改变位点的研究可为水貂阿留申病的致病机理及水貂阿留申病基因疫苗的构建积累资料。

The epidemiological surey showed that Aleutian mink disease is one of the three epideminc disea-ses to the mink farming industry.In spite of several attempts to provide an efficient protective protein based vaccine,experiment have failed so far.Based on published ADV sequences in GenBank,the 12 pairs of primers were designed for ADV in whole gene amplification and sequencing.The sequencing results were compared with the reference strains of ADV and the parvovirus subfamily representative species of each genus for nucleotide sequence and homology analysis.Whole genome amplification and sequencing re-vealed that the sequence lengths of the five strains were 4 537,4 539,4 562,4 572 and 4 569 bp,respec-tively.Nucleotide homology between the five isolated ADV strains and the European or American strains (ADV-G,ADV-Utahl,ADV-SL3)ranged from 92.8% to 97.6%.Based on the amino acid sequence de-duced from VP2 nucleotide of five isolates with DNASTAR software,we found that 61 amino acids were mutated.Three of the strains identified (ADV-G,ADV-DL124 and ADV-DL125)had a nine amino acid deletion at amino acid positions 26-35 and a one amino acid deletion at position 36 (glycine).The changes and deletions of amino acids did not affect the pathogenicity of the ADV.It was the foundation for the study of the Aleutian mink disease pathogenesis and Aleutian mink disease gene vaccine.