Abstract
The baculovirus expression system (BES) is considered to be a very powerful tool for the expression of numerous difficult to express vertebrate proteins. Ssp DnaB mini-intein is a useful fusion partner for the production of recombinant proteins because it can be self-cleaved by controlling the pH and temperature, without additional treatment. To evaluate the utility of Ssp DnaB mini-intein in the BES, recombinant viruses were generated to express the enhanced green fluorescent protein, the VP2 protein of porcine parvovirus, and the E2 protein of classical swine fever virus fused to a mini-intein. As expected, intracellular self-cleavage of the mini-intein occurred during virus infection, but the cleavage initiation time varied depending on the target protein. Significantly enhanced protein production was observed for all of the target proteins that were fused to the mini-intein. This increase was enough to overcome the decrease in the fusion protein due to intracellular self-cleavage. The mini-intein in all of the recombinant fusion proteins was successfully cleaved by controlling the pH and temperature. These results suggest that the Ssp DnaB mini-intein is a useful fusion partner in the BES for easy purification and enhanced production of target proteins.
Highlights
The baculovirus expression system (BES) is widely used for the production of vertebrate proteins or vaccines in insect cells or larvae
These results suggest that the Ssp DnaB mini-intein is a useful fusion partner in the BES for easy purification and enhanced production of target proteins
Bombyx mori 5 (Bm5) cells were cultured at 27 ◦ C and pH 6.4 in TC-100 insect medium (WelGENE, Gyeongsan, GB, Korea) that was supplemented with 10% fetal bovine serum [18]
Summary
The baculovirus expression system (BES) is widely used for the production of vertebrate proteins or vaccines in insect cells or larvae. The fusion partner is frequently highly expressed in host cells, which enhances the expression level of the fusion protein, and favors the purification of the fusion protein [5] These methods have not been applied to all proteins; it is necessary to remove the fusion partner to prevent structural and functional changes to the target protein [6,7]. The target protein can be separated from the fusion partner or an affinity tag without an additional protease treatment [14] This system is an efficacious tool for protein engineering, such as protein expression and purification, because of its self-cleavage capability [11,15,16]. In this study, we applied the Ssp dnaB mini-intein to the baculovirus expression system and reported the optimal conditions for its use and the additional effects of the Ssp dnaB mini-intein
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