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Roles of silkworm endoplasmic reticulum chaperones in the secretion of recombinant proteins expressed by baculovirus system

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Abstract

Baculovirus expression vector system (BEVS) is widely used for production of recombinant eukaryotic proteins in insect larvae or cultured cells. BEVS has advantages over bacterial expression system in producing post-translationally modified secreted proteins. However, for some unknown reason, it is very difficult for insects to secrete sufficiently for certain proteins of interest. To understand the reasons why insect cells fail to secrete some kinds of recombinant proteins, we here employed three mammalian proteins as targets, EPO, HGF, and Wnt3A, with different secretion levels in BEVS and investigated their mRNA transcriptions from the viral genome, subcellular localizations, and interactions with silkworm ER chaperones. Moreover, we observed that no significantly influence on the secretion amounts of all three proteins when depleting or overexpressing most endogenous ER chaperone genes in cultured silkworm cells. However, among all detected ER chaperones, the depletion of BiP severely decreased the recombinant protein secretion in BEVS, indicating the possible central role of Bip in silkworm secretion pathway.

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Acknowledgments

The cost of publication was supported in part by the Research Grant for Young Investigators of Faculty of Agriculture, Kyushu University.

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Author notes

  1. Zhiqing Li

    Present address: State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, P.R. China

Authors and Affiliations

  1. Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, Fukuoka, 812-8581, Japan

    Saki Imai, Takahiro Kusakabe, Jian Xu, Zhiqing Li, Shintaro Shirai, Hiroaki Mon, Daisuke Morokuma & Jae Man Lee

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  1. Saki Imai
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  2. Takahiro Kusakabe
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Correspondence to Jae Man Lee.

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Imai, S., Kusakabe, T., Xu, J. et al. Roles of silkworm endoplasmic reticulum chaperones in the secretion of recombinant proteins expressed by baculovirus system. Mol Cell Biochem 409, 255–262 (2015). https://doi.org/10.1007/s11010-015-2529-5

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