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Ｅ１区缺失的腺病毒载体插入人γ干扰素ｃＤＮＡ序列，与ｐＪＭ１７质粒通过磷酸钙沉淀法共转染２９３细胞，经同源重组，共获得６个病毒噬斑。经ＰＣＲ共扩增检测，证实这６个噬斑均为带有人γ干扰素ｃＤＮＡ的重组腺病毒；受其感染的２９３细胞的上清中，都能测到γ干扰素的活性，且这种活性可被一定稀释度的兔抗人γ干扰素多克隆抗体所中和。其中的１号重组病毒纯化后，按不同的ＭＯＩ（Ｍｕｌｔｉｐｌｅｏｆｉｎｆｅｃｔｉｏｎ）值感染复制缺陷型腺病毒不能在其中增殖的Ｂ１６Ｆ１０细胞，未出现细胞病变效应（Ｃｙｔｏｐａｔｈｉｃｅｆｅｃｔ，ＣＰＥ），而上清中的γ干扰素活性却随ＭＯＩ值增大而升高，这证实构建的复制缺陷型腺病毒是能表达并分泌人γ干扰素的
E1 adenovirus vector was inserted into human interferon-γ cDNA sequence, and pJM17 plasmid co-transfected 293 cells by calcium phosphate precipitation, homologous recombination, a total of 6 virus plaques. By PCR co-amplification test confirmed that these six plaques are with human γ interferon cDNA recombinant adenovirus; 293 cells infected by its infection in the supernatant, we can detect γ interferon activity, and This activity can be neutralized by a dilution of rabbit anti-human interferon gamma polyclonal antibody. After purification of the recombinant virus no. 1, B16F10 cells which replication-defective adenovirus could not proliferate were infected by different MOI (Multipleofinfection), no cytopathic effect (CPE) was observed, but γ-interference in the supernatant Superoxide dismutase activity increased with increasing MOI, which confirmed that the constructed replication-deficient adenovirus was able to express and secrete human IFN-γ