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Enhanced Protein Secretion From Insect Cells by Co-Expression of the Chaperone Calreticulin and Translation Initiation Factor eIF4E

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Abstract

Host protein synthesis is shut down in the lytic baculovirus expression vector system (BEVS). This also affects host proteins involved in routing secretory proteins through the endoplasmic reticulum (ER)-Golgi system. It has been demonstrated that a secretory alkaline phosphatase–EGFP fusion protein (SEFP) can act as a traceable and sensitive secretory reporter protein in BEVS. In this study, a chaperone, calreticulin (CALR), and the translation initiation factor eIF4E were co-expressed with SEFP using a bicistronic baculovirus expression vector. We observed that the intracellular distribution of SEFP in cells co-expressing CALR was different from co-expressing eIF4E. The increased green fluorescence emitted by cells co-expressing CALR had a good correlation with the abundance of intracellular SEFP protein and an unconventional ER expansion. Cells co-expressing eIF4E, on the other hand, showed an increase in extracellular SEAP activity compared to the control. Utilization of these baculovirus expression constructs containing either eIF4E or CALR offers a significant advantage for producing secreted proteins for various biotechnological and therapeutic applications.

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Abbreviations

BEVS:

Baculovirus expression vector system

IRES:

Internal ribosome entry site

SEAP:

Secreted alkaline phosphatase

SEFP:

SEAP–EGFP fusion protein

ER:

Endoplasmic reticulum

Sf21:

Spodoptera frugiperda 21

MOI:

Multiplicity of infection

CALR:

Calreticulin

eIF4E:

Eukaryotic initiation factor 4E

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Authors and Affiliations

  1. Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan

    Chao-Yi Teng & Shou-Lin Chang

  2. Laboratory of Virology, Wageningen University, Wageningen, The Netherlands

    Chao-Yi Teng & Monique M. van Oers

  3. Department of Bioscience Technology, Chung Yuan Christian University, Chungli, Taiwan

    Tzong-Yuan Wu

  4. R&D Center of Membrane Technology, Chung Yuan Christian University, Chungli, Taiwan

    Tzong-Yuan Wu

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  1. Chao-Yi Teng
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  2. Shou-Lin Chang
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  4. Tzong-Yuan Wu
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Correspondence to Tzong-Yuan Wu.

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Teng, CY., Chang, SL., van Oers, M.M. et al. Enhanced Protein Secretion From Insect Cells by Co-Expression of the Chaperone Calreticulin and Translation Initiation Factor eIF4E. Mol Biotechnol 54, 68–78 (2013). https://doi.org/10.1007/s12033-012-9545-4

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