For the development of mink Aleutian disease nucleic acid vaccine,the nucleotide sequences encoding 428 ~446 bit amino acids in the ADV VP2 gene are removed by using the overlap extension PCR technique,with pcDNA3.1 (+)carrier connection,construct the whole gene mutation recombinant plasmid pcDNA3.1 -ADV -428,on this basis,nucleotide sequences of 487 ~501 amino acid at position is truncated coding,construct the whole gene mutation recombinant plasmid pcDNA3.1 -ADV -428 -487.The recombinant plasmid was constructed by intramuscular injection of immune mice,anti ADV antibody levels were detected in 14 days,28 days,42 days,and 56 days after inoculation by indirect ELISA method;Detection of CD3 +,CD4 + and CD8 +T lymphocyte subsets in spleen cells of mice forty -second days after inoculation by flow cytometry.Results showed that the number of CD3 +,CD4 + and CD8 + T lymphocyte subsets in mice inoculated with plasmid were significantly increased,anti ADV antibody level reached peak at forty -second days.In this experiment,the immunogenicity of the recombinant plasmid of ADV gene mutation was tested to provide reference for the development of mink Aleutian disease nucleic acid vaccine.%为研制水貂阿留申病核酸疫苗，应用重叠延伸 PCR 技术去除 ADV VP2基因中编码428～446位氨基酸的核苷酸序列，与 pcDNA3．1（＋）载体连接，构建全基因突变重组质粒 pcDNA3．1－ADV －428，在此基础上，截去编码487～501位氨基酸的核苷酸序列，构建全基因突变重组质粒pcDNA3．1－ADV －428－487。将构建的重组质粒经肌肉注射免疫小鼠，应用间接 ELISA 法检测接种后14、28、42、56 d 抗 ADV 抗体水平；流式细胞术检测接种后第42天小鼠脾细胞 CD3＋、CD4＋和CD8＋T 淋巴细胞亚群。结果显示，小鼠接种质粒后 CD3＋、CD4＋和 CD8＋T 淋巴细胞亚群数量均明显增加，第42天抗 ADV 抗体水平达峰值。本试验通过对 ADV 全基因突变重组质粒的免疫原性进行分析，为水貂阿留申病核酸疫苗的研制提供了参考。
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