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将ＡｃＮＰＶＤＮＡ和ＢａｍＨＩ酶解的ＢｍＮＰＶＤＮＡ共转染ＳＦ２１细胞，再用子代病毒反复感染ＳＦ２１和ＢｍＮ细胞。经空斑分析纯化，筛选出既能在ＳＦ细胞又能在ＢｍＮ细胞中复制多角体病毒的昆虫核型多角体病毒。限制酶分析表明筛选所得的病毒为ＡｃＮＰＶ和ＢｍＮＰＶ的杂交型（ＨｙＮＰＶ）。进一步应用本室构建的携有木质素酶Ｈ８同功酶基因的ＳＦ细胞表达载体ｐＶＬ－Ｍｕ－Ｈ８与ＨｙＮＰＶ共转染ＳＦ细胞，纯化筛选获得拓宽宿主范围的重组病毒（ｐＶＬ－Ｍｕ－Ｈ８ＨｙＮＰＶ）。ＥＣＬＤｏｔ－ｂｌｏｔＤＮＡ杂交分析显示用ｐＶＬ－Ｍｕ－Ｈ８ＨｙＮＰＶ感染ＳＦ和ＢｍＮ细胞的总ＤＮＡ中均含有与木质素酶Ｈ８同功酶基因ｃＤＮＡ（λＭＬ－１ＤＮＡ）探针杂交的ＤＮＡ片段。感染细胞裂解液的Ｗｅｓｔｅｒｎｂｌｏｔ分析证明该重组病毒既能在ＳＦ细胞也能在ＢｍＮ细胞中异源表达外源基因—木质素酶Ｈ８同功酶，呈现完全相同的Ｗｅｓｔｅｒｎｂｌｏｔ分析图谱。并将表达产物分泌到胞外。
SF21 cells were co-transfected with AcNPVDNA and BamHI-digested BmNPVDNA, and SF21 and BmN cells were repeatedly infected with progeny virus. After the plaque analysis and purification, we screened the insect nuclear polyhedrosis virus that can replicate polyhedrosis virus in both SF cells and BmN cells. Restriction enzyme analysis indicated that the selected viruses were hybrids (HyNPV) of AcNPV and BmNPV. Further, SF cell expression vector pVL-Mu-H8 and HyNPV carrying ligninase H8 isoenzyme gene constructed in our laboratory were co-transfected into SF cells, and the host cell range of the recombinant virus (pVL-Mu-H8HyNPV) . ECLDot-blot DNA hybridization analysis showed that the total DNA of SF and BmN cells infected with pVL-Mu-H8HyNPV contained DNA fragments hybridized with the cDNA of the ligninase H8 isozyme gene (λML-1 DNA). Western blot analysis of the infected cell lysate demonstrated that the recombinant virus heterologously expressed the exogenous gene ligninase H8 isoenzyme in SF cells as well as in BmN cells and showed identical Western blot analysis. And secreted the expression product to the extracellular.