title: Efficient protein production using a Bombyx mori nuclear polyhedrosis virus lacking the cysteine proteinase gene.
abstract: Infection by a baculovirus (Bombyx mori nuclear polyhedrosis virus, BmNPV) in silkworm (Bombyx mori) larvae is highly efficient as an expression system for the production of useful proteins. However, the amount of the protein of interest expressed tends to decrease in the later stages of infection presumably due, in part, to a proteinase produced in the larval haemolymph. The N-terminal amino acid sequence of a proteinase purified from the haemolymph of BmNPV-infected larvae was identical to the internal amino acid sequence of the viral cysteine proteinase gene of BmNPV, suggesting that the cysteine proteinase in the haemolymph originated from the BmNPV gene. We constructed a mutant virus (CPd) which had a deletion in the cysteine proteinase gene. No proteinase activity corresponding to this proteinase was detected in the haemolymph of silkworm larvae infected with CPd. The firefly luciferase and the human growth hormone genes were separately introduced into CPd under control of the polyhedrin promoter. These constructs produced these proteins very efficiently, because of a greatly reduced degree of degradation of these proteins. A BmNPV vector system using CPd enhances the stability of foreign expressed proteins, especially for those that are cysteine proteinase-sensitive.
authors.affiliation: Research Institute for Biological Science, Katakura Industries Co., Ltd, Matsumoto, Nagano, Japan. ribs@po.cnet.ne.jp
authors.forename: T
authors.initials: T
authors.lastname: Suzuki
authors.forename: T
authors.initials: T
authors.lastname: Kanaya
authors.forename: H
authors.initials: H
authors.lastname: Okazaki
authors.forename: K
authors.initials: K
authors.lastname: Ogawa
authors.forename: A
authors.initials: A
authors.lastname: Usami
authors.forename: H
authors.initials: H
authors.lastname: Watanabe
authors.forename: K
authors.initials: K
authors.lastname: Kadono-Okuda
authors.forename: M
authors.initials: M
authors.lastname: Yamakawa
authors.forename: H
authors.initials: H
authors.lastname: Sato
authors.forename: H
authors.initials: H
authors.lastname: Mori
authors.forename: S
authors.initials: S
authors.lastname: Takahashi
authors.forename: K
authors.initials: K
authors.lastname: Oda
mesh_terms: D000818:Animals; D012831:Bombyx / Q000821:virology*; D003546:Cysteine Endopeptidases / Q000235:genetics*; D017353:Gene Deletion; D006801:Humans; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000378:metabolism; D014176:Protein Biosynthesis
chemical_list: D003546:Cysteine Endopeptidases