中国生物工程杂志

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.