AI Article Synopsis
- The baculovirus expression system (BES) is effective for producing tough vertebrate proteins, and Ssp DnaB mini-intein serves as a beneficial fusion partner that can self-cleave under controlled conditions.
- Recombinant viruses were created to express multiple proteins fused to the mini-intein, resulting in successful intracellular cleavage during infection, although the timing varied with each protein.
- Overall, using Ssp DnaB mini-intein significantly boosted protein production, helping to counterbalance the loss from self-cleavage, and demonstrates its potential for easier purification and enhanced yields in BES applications.
Article 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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6213604 | PMC |
| http://dx.doi.org/10.3390/v10100523 | DOI Listing |
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