期刊
MOLECULAR BIOTECHNOLOGY
卷 61, 期 11, 页码 852-859出版社
HUMANA PRESS INC
DOI: 10.1007/s12033-019-00210-5
关键词
Bacmid; Capsid; Dengue virus; Premembrane; Envelope; Silkworm
资金
- JSPS KAKENHI [16K14900]
- Research Program for Next Generation Young Scientists of Network Joint Research Center for Materials and Devices: Dynamic Alliance for Open Innovation Bridging Human, Environment and Materials
- Grants-in-Aid for Scientific Research [16K14900] Funding Source: KAKEN
To explore virus-like particles formation of dengue virus serotype type 2 (DENV-2) structural proteins of, C, prM, E were expressed in silkworm larvae using recombinant Bombyx mori nucleopolyhedroviruses (BmNPV). Each recombinant BmNPV bacmid coding the 2C-prM-E polypeptide and E protein fused with the signal peptide of bombyxin from B. mori was injected into silkworm larvae. The expressed proteins were collected from hemolymph and fat body, and purified using affinity chromatography. E protein was observed at 55 kDa. The DENV virus-like particles (DENV-LPs) with a diameter approximately 35 nm was observed using transmission electron microscopy (TEM) and immunogold-labelling TEM analysis. The binding of each partially purified proteins to heparin, one of receptors for DENV was confirmed. DENV-LPs were secreted in silkworm larval hemolymph even still low amount, but the E protein and heparin binding function were confirmed.
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