Optimal Conditions for the Expression of Glycoprotein E2 of Classical Swine Fever Virus using Baculovirus in Insect Cells
The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein Erns (E0), E1 and E2. Among them, E2, the most immunogenicof the CSFV glycoproteins, induces a protective immune response in swine. In this study, todetermine the optimal express...
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Veröffentlicht in: | International Journal of Industrial Entomology 2014, 29(2), , pp.207-213 |
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Zusammenfassung: | The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein Erns (E0), E1 and E2. Among them, E2, the most immunogenicof the CSFV glycoproteins, induces a protective immune response in swine. In this study, todetermine the optimal expression conditions of glycoprotein E2 using baculovirus system,we investigated the influence of insect cells and media to the expression of recombinant E2.
Recombinant virus containing glycoprotein E2 coding gene was constructed with bApGOZADNA. Expression of the glycoprotein E2 was analyzed by SDS-PAGE and Western blotanalysis using anti-CSFV E2 monoclonal antibodies. Expression of glycoprotein E2 in Sf21cells was first observed after 3 days and reached a maximum on the 5th day after infection.
Furthermore, the highest levels of glycoprotein E2 expression were observed at multiplicity ofinfection (MOI) of 5. When three different insect cell lines (Sf21, High-Five and Se301) weretested, High-Five cells showed the highest production. In addition, four different serum-freeand serum-supplemented media, respectively, were tested for the expression of glycoproteinE2 and the budded virus (BV) titers. As a result, serum-supplemented medium provided thebest conditions for protein production and the BV yield. KCI Citation Count: 0 |
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ISSN: | 1598-3579 2586-4785 |
DOI: | 10.7852/ijie.2014.29.2.207 |