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- W2386501268 abstract "Objective To express hepatitis E virus structural protein p166 (HEVp166) in Silkworm/Bombyx mori nucleopolyhedrovirus (BmNPV) expression vector system. Methods Gene fragment of HEVp166 was amplified by polymerase chain reaction (PCR) and then inserted into the downstream of polyhedron promoter of the BmNPV genome. Recombinant virus was constructed by gene cloning, co-transfection, and plaque purification. Then the HEVp166 was expressed in BmN cells and silkworm pupae. The expression of HEVp166 in BmN cell (harvested at 36 h postinfection) was detected by indirect immunofluorescence assay (IIFA), and the expression of HEVp166 in silkworm pupa (harvested at 144 h postinfection) was detected by SDS-PAGE and western blotting. Results The recombinant baculovirus expression vector Bm-HEV166 containing HEVp166 was constructed successfully. The p166 protein was expressed in BmN cells and detected as green globoids by IIFA. The p166 protein was efficiently expressed in silkworm pupae and obtained from pupal hemolymph of the pupae infected with the recombinant virus Bm-HEV166. The expressed protein combined specifically with monoclonal antibody of anti-p166 and formed a protein band of about 19 kD in the western blotting profiles. Conclusion The HEV p166 could be expressed at higher level in silkworm pupae with Silkworm/Bm-(NPV) expression vector system, and might be used as a candidate for studying oral vaccine of HEV." @default.
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- W2386501268 date "2005-01-01" @default.
- W2386501268 modified "2023-09-25" @default.
- W2386501268 title "Expression of hepatitis E virus structural protein p166 gene fragment in BmN cell and silkworm pupa" @default.
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