柞蚕核型多角体病毒载体在培养细胞和休眠蛹中的基因表达效果

柞蚕核型多角体病毒（AnpeNPV）作为基因表达载体在柞蚕培养细胞（AnPe细胞）和柞蚕蛹中已经成功地表达出了外来基因，并生产出了大量蛋白质。本文比较了AnpeNPV与苜蓿尺蠖核型多角体病毒(AcMNPV)、家蚕核型多角体病毒(BmNPV)和美国白蛾核型多角体病毒(HycuNPV)基因表达载体在培养细胞和昆虫活体组织内的β-半乳糖苷酶基因表达效果。结果显示，5×105个细胞中β-半乳糖苷酶的最高酶活性分别是AnpeNPV在AnPe细胞为40.9 units/ml （TC-100培养液，FBS10%）和59.9 units/ml（SF-900Ⅱ培养液），AcMNPV在Sf9细胞为72.4 units/ml（TC-100，FBS10%）和66.4 units/ml（SF-900Ⅱ）、在High5细胞为326 units/ml（EX-CELL 405培养液），BmNPV在Bm4细胞为15.1 units/ml（TC-100，FBS10%），HycuNPV在SpIm细胞为68.6 units/ml（SF-900Ⅱ）。活体组织内β-半乳糖苷酶的最高酶活性分别是柞蚕雌蛹为14.3 units/g、雄蛹为11.7 units/g，家蚕幼虫是10.1 units/g。实验证明AnpeNPV/AnPe的外来基因表达水平与AcMNPV/ Sf9和HycuNPV/SpIm相似、比BmNPV/ Bm4高、不及AcMNPV/ High5；AnpeNPV/柞蚕蛹，其雌蛹比BmNPV/家蚕5龄幼虫的外来基因表达效果好、雄蛹与之无明显差异，说明AnpeNPV基因表达载体无论是在培养细胞还是昆虫活体组织中均可与其他NPV基因表达载体相媲美。柞蚕蛹由于可以机械化、大规模地操作，显示对于大量生产蛋白质具有更好的应用前景。

Gene Expression Efficiency of Antheraea Pernyi Nucleopolyhedrovirus Vector in Both an Established Cell Line and Diapausing Pupae

Chinese oak silkworm, Antheraea pernyi nuclear polyhedrosis virus (AnpeNPV) as a gene expression vector using A. pernyi culture cell (AnPe cell ) and diapausing pupa has been successfully expressed a foreign gene and produced a great lot protein. This paper compared the expression efficiency of β-galactosidase gene between AnpeNPV with AcMNPV, BmNPV and HycuNPV gene expression vectors in insect culture cells and live insect tissues. The experiments results showed that the maximum β-galactosidase activity were AnpeNPV cells 40.9 units/ml (TC-100 medium, FBS 10%) and 59.9 units/ml (SF-900 medium II) in AnPe cells, AcMNPV 72.4 units/ml (TC-100, FBS 10%) and 66.4 units/ml (SF-900 II) in Sf9 cells, 326 units/ml in High5 cells (EX-405 medium), BmNPV 15.1 units/ml in Bm4 cells (TC-100, FBS 10%), HycuNPV 68.6 units/ml in SpIm cells (SF-900 II) in 5 × 105 cells, individually. In live insect tissues, the maximum β- galactosidase activity were A. pernyi female pupae 14.3 units/g, male pupae 11.7 units/g, Bombyx mori 5th instar larvae 10.1 units/g. In culture cells, the β-galactosidase level of AnpeNPV was similar to AcMNPV with exception of using High5 cells and to HycuNPV, better to BmNPV. In live insect tissues, the β-galactosidase level of AnpeNPV was similar (male pupa) or significantly higher (female pupa) than BmNPV. AnpeNPV gene expression vector exhibited similar expression efficiency to other NPV vectors in both cultured cells or live insect tissue. Compared to other expression system. The diapausing pupa is easily operated, and suitable for large amout production.