中国生物工程杂志

Chinese oak silkworm, Antheraea pernyi is an economic insect rearing outdoors. It is mainly cultured in the area of Northeast of China. A. pernyi diapause by pupa stage for hibernation. The pupa is big, easy fix, keep time long, need not the feeding, easy conveyance etc. advantage. A. pernyi pupae as biological reactor product foreign protein may be large-scale mechanization production, reduce operational and trivial with labor force. In this paper, A. pernyi nucleopolyhedrovirus (AnpeNPV) as gene expression vector successfully expressed β–galatosidase gene (LacZ) in A. pernyi culture cells (AnPe cell) and diapause pupae, moreover purified recombinant AnpeNPV expressing LacZ (AnpeLacZ) obtained using AnPe cell. For β–galatosidase yield of the AnpeLacZ, their peak activity was 40.9 units/ml at 12 days post-infection (p. i.) in AnPe cell cultured using TC-100 medium (10 % FBS) and is 59.9 units/ml at 18 days p. i. in AnPe cell cultured using SF-900Ⅱ medium, respectively. The latter was slightly high but slow at time. The β–galatosidase activity of AnpeLacZ in A. pernyi pupae who store 7 months at 5 ℃ reached peak at 15 days p. i., female was 14.3 units/g and male was 11.7 units/g, respectively. The female was better than male. Results demonstrate that AnpeNPV and A. pernyi pupae could be developed and used as a novel baculovirus gene expression system which may be mechanization and large-scale production.