期刊
SPRINGERPLUS
卷 1, 期 -, 页码 -出版社
SPRINGER INTERNATIONAL PUBLISHING AG
DOI: 10.1186/2193-1801-1-29
关键词
Virus-like particles; Human papillomavirus silkworm expression system; BmNPV bacmid
资金
- Promotion of Nanobio-Technology Research
- Ministry of Education, Culture, Sports, Science & Technology (MEXT), Japan
- Japanese Government Scholarship fellow MEXT, Japan
Human papillomavirus (HPV) 6b L1 capsid protein was expressed using the Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid expression system in silkworm larvae. Two constructs, full-length L1 (500 a.a) and C-terminal-deleted short L1 (479 a.a), and three PCR-manipulated antigenic loops at amino acids 55-56, 174-175, and 348-349 regions were incorporated with whole enhanced green fluorescent protein (EGFP). Expressed in full, short L1 proteins and variants were purified in heparin affinity column chromatography and confirmed by SDS-PAGE and western blot. The presence of self-assembled virus-like particles (VLPs) and EGFP incorporation on the surface of VLPs were confirmed by the observation of transmission electron and immunoelectron microscopies, respectively. HPV 6b L1 major capsid protein was successfully expressed in silkworm, and effective manipulation on the antigenic regions showed the path to versatile vaccine development based on HPV L1-VLPs.
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