Construction of a cysteine protease deficient Bombyx mori multiple nucleopolyhedrovirus bacmid and its application to improve expression of a fusion protein
Copyright policy )Construction of a cysteine protease deficient Bombyx mori multiple nucleopolyhedrovirus bacmid and its application to improve expression of a fusion protein
The bacmid system of BmMNPV with cysteine protease gene deletion (CPD-BmMNPV bacmid) was constructed using the lambda recombination system. The protease activities of Bombyx mori cells and silkworm larvae infected with this CPD-BmMNPV bacmid were reduced by 94% and 85%, respectively. By using this system, a GFP(uv)-beta1,3-N-acetylglucosaminyltransferase 2 (GFP(uv)-beta3GnT2) fusion protein was successfully expressed in silkworm larvae with less protein degradation and without larvae liquefaction; beta3GnT activity improved 30%. This CPD-BmMNPV bacmid system provides rapid protein production in silkworms and can be used for the production of recombinant eukaryotic proteins without proteolytic degradation.
- Shizuoka University Japan
Base Sequence, Recombinant Fusion Proteins, Genetic Vectors, Molecular Sequence Data, Gene Expression, Bombyx, N-Acetylglucosaminyltransferases, Protein Engineering, Nucleopolyhedroviruses, Cell Line, Cysteine Endopeptidases, Larva, Animals, Cloning, Molecular
Base Sequence, Recombinant Fusion Proteins, Genetic Vectors, Molecular Sequence Data, Gene Expression, Bombyx, N-Acetylglucosaminyltransferases, Protein Engineering, Nucleopolyhedroviruses, Cell Line, Cysteine Endopeptidases, Larva, Animals, Cloning, Molecular
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