阿留申病毒大连株非结构蛋白基因的克隆与序列分析

Cloning and Sequencing of Non-structural Genes of Aleutian Mink Disease Virus of Dalian Strains

根据Genbank上发表的水貂阿留申病毒基因序列数据，设计了3对引物，采用PCR的方法分别对ADV—DL1、ADV—DL2株非结构蛋白基因进行扩增，将各片断克隆至pMD18-T载体，经PCR鉴定后进行了序列测定和分析。结果显示，ADV—DL1、ADV—DL2与GenBank上公布的ADV毒株相比，核苷酸序列同源率NS1为85．9％-90．0％．Ns2为85．1％-92．7％．NS3为83．0％～95．1％。ADV—DL1与ADV—DL2同源率NS1为93．7％，NS2为95．6％，NS3为98．5％。氨基酸同源率NS1为82．8％-85．8％，NS2为80．7％～92．1％，Ns3为72．9％～95．4％。ADV—DL1与ADV—DL2同源率NS1为90．3％，NS2为92．1％，NS3为95．4％。

Based on the sequences of the non-structural （NS） gene of Aleutian mink disease virus （ADV） published in Genbank, three pairs of primer were designed to amplify the NS genes of ADV-DL1 and ADV-DL2 strains. The NS gene was amplified by polymerase chain reaction （PCR）, and subsequently cloned into pMD18-Tvector. Results of sequencing analysis showed that the nucleotide homologies between ADV-DL1, ADV-DL2 and published sequences were 85.9% - 90.0% for NS1, 85.1% -92.7% for NS2, 83.0% -95.1% for NS3, and those between ADV-DL1 and ADV-DL2 were 93.7%, 95.6% and 98.5% for NS1, NS2 and NS3, respectively. The homological rates of amino acid was 82.8% -85.8% for NS1, 80.7% - 92. 1% for NS2, and 72.9% - 95.4% for NS3, and those of ADV-DL1 and ADV-DL2 was 90.3 %, 92.1% and 95.4% for NS1, NS2 and NS3, respectively.