Analysis of time dependent expression of recombinant Crimean-Congo hemorrhagic fever virus Gn protein in insect cells

The Crimean-Congo hemorrhagic fever virus (CCHFV) belongs to family Bunyaviridea, genus Nairovirus. This virusencodesfour structural proteins nucleoprotein (N), amino terminal glycoprotein (Gn), carboxyterminal glycoprotein (Gc), and L protein, 78-kD. Using the baculovirusexpressionsystem, we expressed the full-length coding sequence of the Gn glycoprotein optimized for expression in Sf9 cells and analyzed its expression in different times in order to find best time of cell harvesting for further analysis. SDS-PAGE analysis of cell cruds showed that time between 36-48 hours post infection was the best time for recombinant antigen harvesting to obtain high amount of recombinant proteinand delay may result in losing of recombinant antigen. This knowledge could be valuable for further experiments like extraction of Gn from Sf9 cells for purification or immunization assay that needs high amount of recombinant protein in initial samples.

Keywords:

Crimean-Congo hemorrhagic fever virus , glycoprotein , Gn , baculovirus , Sf9 cell