alexa Improved expression of fusion protein using a cysteine- protease- and chitinase-deficient Bombyx mori (silkworm) multiple nucleopolyhedrovirus bacmid in silkworm larvae.
Pharmaceutical Sciences

Pharmaceutical Sciences

Journal of Bioequivalence & Bioavailability

Author(s): Park EY, Abe T, Kato T

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Abstract A bacmid system of BmMNPV [Bombyx mori (silkworm) multiple nucleopolyhedrovirus] with both the cysteine-protease and chitinase genes deleted (BmMNPV-CP(-)-Chi(-)) was constructed. The viral protease and chitinase activities in the haemolymph of Bombyx mori larvae infected with this BmMNPV-CP(-)-Chi(-) bacmid were reduced by 95 and 50\% respectively. By using this system, a GFP(uv)-beta3GnT2 (green fluorescent protein excited by UV light-beta1,3-N-acetylglucosaminyltransferase 2) fusion protein was successfully expressed in silkworm larvae with less protein degradation and without larvae liquefaction; beta3GnT2 activity improved 2.8-fold over that of unmodified bacmid. This BmMNPV-CP(-)-Chi(-) bacmid system provides rapid protein production in silkworms and can be used for the production of recombinant eukaryotic proteins without proteolytic degradation. This article was published in Biotechnol Appl Biochem and referenced in Journal of Bioequivalence & Bioavailability

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