RECOMBINANT BOMBYX MORI NUCLEAR POLYHEDROSIS VIRUS LACKING THE VIRUS-ENCODED CHITINASE OR CATHEPSIN GENES

摘要

A recombinant bombyx mori nuclear polyhedrosis virus in which virus-encoded chitinase or cathepsin genes are deleted is provided to be usefully used for mass-producing foreign proteins in bombyx mori. A cathepsin(v-cath) gene of a recombinant bombyx mori nuclear polyhedrosis virus(RecBmNPV-v-cath^-) is deleted and chitinase(chiA) gene of a recombinant bombyx mori nuclear polyhedrosis virus(RecBmNPV-chiA^-) is deleted. Both cathepsin(v-cath) and chitinase(chiA) of a recombinant bombyx mori nuclear polyhedrosis virus(RecBmNPV-chiA^--v-cath^-) are deleted. A method for preparing the recombinant bombyx mori nuclear polyhedrosis virus(RecBmNPV-chiA^--v-cath^-) comprises the steps of: (a) preparing a recombinant bombyx mori nuclear polyhedrosis virus(RecBmNPV) expressing cellulase gene; (b) amplifying a 4.3kb DNA fragment including cathepsin(v-cath) and chitinase(chiA) from the RecBmNPV through PCR and cloning the PCR product into pGemT vector to prepare a transfer vector(pGemT-chiA-v-cath); (c) treating the v-cath in the transfer vector with Xbal and Xhol to be replaced with a GFP gene and treating the chiA gene in the vector with KpnI and SmaI to be replaced with a Luci gene to prepare a recombinant transfer vector(pGemT-Luci-GFT); and (d) co-transfecting the recombinant transfer vector(pGemT-Luci-GFT) and the recombinant virus(RecBmNPV) into Bm5 to prepare a recombinant virus in which both the v-cath and chiA are deleted.