caa a22 4500 606234853 CHVBK 20210128101239.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s11010-015-2529-5 doi (NATIONALLICENCE)springer-10.1007/s11010-015-2529-5 Roles of silkworm endoplasmic reticulum chaperones in the secretion of recombinant proteins expressed by baculovirus system [Elektronische Daten] [Saki Imai, Takahiro Kusakabe, Jian Xu, Zhiqing Li, Shintaro Shirai, Hiroaki Mon, Daisuke Morokuma, Jae Lee] 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. Springer Science+Business Media New York, 2015 Baculovirus expression system nationallicence Recombinant protein nationallicence RNAi nationallicence ER chaperone nationallicence Silkworm nationallicence Imai Saki Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Kusakabe Takahiro Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Xu Jian Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Li Zhiqing Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Shirai Shintaro Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Mon Hiroaki Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Morokuma Daisuke Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Lee Jae Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 409/1-2(2015-11-01), 255-262 0300-8177 409:1-2<255 2015 409 11010 https://doi.org/10.1007/s11010-015-2529-5 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2529-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Imai Saki Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 700 1- Kusakabe Takahiro Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 700 1- Xu Jian Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 700 1- Li Zhiqing Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 700 1- Shirai Shintaro Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 700 1- Mon Hiroaki Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 700 1- Morokuma Daisuke Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 700 1- Lee Jae Laboratory of Insect Genome Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, 812-8581, Fukuoka, Japan aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 409/1-2(2015-11-01), 255-262 0300-8177 409:1-2<255 2015 409 11010