Enhanced Production of Recombinant Protein by Fusion Expression with Ssp DnaB Mini-Intein in the Baculovirus Expression System
| Autor: | Sung Min Bae, Beom Ku Han, Soo Dong Woo, Jae Bang Choi, Won Seok Gwak |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Porcine parvovirus purification Recombinant Fusion Proteins Genetic Vectors Green Fluorescent Proteins lcsh:QR1-502 Gene Expression Virus Article lcsh:Microbiology law.invention Cell Line Inteins Ssp DnaB mini-intein 03 medical and health sciences baculovirus Viral Envelope Proteins law Virology Protein biosynthesis Animals Antigens Viral dnaB helicase biology Chemistry Temperature Hydrogen-Ion Concentration biology.organism_classification Bombyx Fusion protein Cell biology 030104 developmental biology Infectious Diseases Recombinant DNA Capsid Proteins Target protein fusion partner Intein Baculoviridae enhanced production |
| Zdroj: | Viruses, Vol 10, Iss 10, p 523 (2018) Viruses Volume 10 Issue 10 |
| ISSN: | 1999-4915 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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