Abstract
BackgroundThere are few studies that have examined the potential of RNA inference (RNAi) to increase protein production in the baculovirus expression vector system (BEVS). Spodoptera frugiperda (fall armyworm) (Sf)-caspase-1-repressed stable cells exhibit resistance to apoptosis and enhancement of recombinant protein production. However, the mechanism of recombinant protein augmentation in baculovirus-infected Caspase-repressed insect cells has not been elucidated.ResultsIn the current study, we utilized RNAi-mediated Sf-caspase-1-repressed stable cells to clarify how the resistance to apoptosis can enhance both intracellular (firefly luciferase) and extracellular (secreted alkaline phosphatase [SEAP]) recombinant protein production in BEVS. Since the expression of molecular chaperones is strongly associated with the maximal production of exogenous proteins in BEVS, the differential expression of molecular chaperones in baculovirus-infected stable cells was also analyzed in this study.ConclusionThe data indicated that the retention of expression of molecular chaperones in baculovirus-infected Sf-caspase-1-repressed stable cells give the higher recombinant protein accumulation.
Highlights
There are few studies that have examined the potential of RNA inference (RNAi) to increase protein production in the baculovirus expression vector system (BEVS)
After more than 20 passages, stable cell lines were analyzed by genomic DNA polymerase chain reaction (PCR) and RT-PCR to examine the inverted-repeat DNA sequence of Sf-caspase-1 and endogenous expressed Sfcaspase-1 mRNA quantity
The data showed that levels of Sf-casp1 mRNA in S. frugiperda 9 (Sf9)/pIBdsCasp-1 and Sf9/ pIBdsCasp-2 cells were apparently lower than those observed in Sf9 and Sf9/pIB cells (Figure 3B)
Summary
There are few studies that have examined the potential of RNA inference (RNAi) to increase protein production in the baculovirus expression vector system (BEVS). Spodoptera frugiperda (fall armyworm) (Sf)-caspase-1repressed stable cells exhibit resistance to apoptosis and enhancement of recombinant protein production. Apoptosis plays an important role during development and tissue homeostasis eliminating discarded cells from the organism, including damaged and virus-infected cells. For this reason, apoptosis acts as a host protection mechanism by which virus-infected cells are removed to limit the proliferation of viruses [4,5]. Apoptosis acts as a host protection mechanism by which virus-infected cells are removed to limit the proliferation of viruses [4,5] The Op-IAP from the Orgyia pseudotsugata multicapsid nucleopolyhedrovirus (OpMNPV) and Sf-IAP from the host, S. frugiperda cells, suppress the apoptosis process (Figure 1) [1,2]
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