Additive effect of calreticulin and translation initiation factor eIF4E on secreted protein production in the baculovirus expression system |
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Authors: | Chao-Yi Teng Monique M van Oers Tzong-Yuan Wu |
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Institution: | 1. Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li, 320, Taiwan, Republic of China 2. Laboratory of Virology, Wageningen University, Wageningen, The Netherlands 3. Center for Nanotechnology and Institute of Biomedical Technology, Chung Yuan Christian University, Chung-Li, 320, Taiwan, Republic of China
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Abstract: | The baculovirus expression vector system is widely used for the production of recombinant proteins. However, the yield of membrane-bound or secreted proteins is relatively low when compared with intracellular or nuclear proteins. In a previous study, we had demonstrated that the co-expression of the human chaperones calreticulin (CALR) or β-synuclein (β-syn) increased the production of a secreted protein considerably. A similar effect was also seen when co-expressing insect translation initiation factor eIF4E. In this study, different combinations of the three genes were tested (CALR alone, β-syn?+?CALR, or β-syn?+?CALR?+?eIF4E) to further improve secretory protein production by assessing the expression level of a recombinant secreted alkaline phosphatase (SEFP). An additional 1.8-fold increment of SEFP production was obtained when cells co-expressed all the three “helper” genes, compared to cells, in which only CALR was co-produced with SEFP. Moreover, the duration of the SEFP production lasted much longer in cells that co-expressed these three “helper” genes, up to 10 dpi was observed. Utilization of this “triple-supporters” containing vector offers significant advantages when producing secreted proteins and is likely to have benefits for the production of viral vaccines and other pharmaceutical products. |
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