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蚕核多角体ウイルスを用いた日本脳炎ウイルス外被膜糖タンパク(E)の発現


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Title: 蚕核多角体ウイルスを用いた日本脳炎ウイルス外被膜糖タンパク(E)の発現
Other Titles: Expression of Envelope Glycoprotein E of Japanese Encephalitis Virus using Bombix mori Nuclear Polyhedrosis Virus
Authors: 姶良, 義一
Authors (alternative): Aira, Yoshikazu
Issue Date: 28-Dec-1987
Publisher: 長崎大学熱帯医学研究所 / Institute of Tropical Medicine, Nagasaki University
Citation: 熱帯医学 Tropical medicine 29(4). p195-209, 1987
Abstract: In order to prevent and control Japanese encephalitis (JE) in presently epidemic areas, development of the second generation JE vaccine using recombinant DNA technology has been postulated by the World Health Organization (WHO). Toward this objective, the author tried to express JE virus envelope glycoprotein (E) using Bombix mori nuclear polyhedrosis virus as a baculovirus expression vector. The cDNA segment approximately corresponding to the entire E protein gene was prepared from E. coli plasmid pUC13 inserted with cDNA fragment S22, which covers entire E protein gene with its flanking sequences on both ends. The E protein gene cDNA was then modified and inserted to the transfer vector plasmid pBF 48 which carries baculovirus polyhedrin gene promotor and terminal regions with their flanking sequences. In the resulting recombinant vector plasmid, the E protein gene should be linked to the polyhedrin gene promotor and the terminator in correct orientation. The DNAs from the recombinant vector plasmid and wild type baculovirus were co-transfected to BmN cells and recombinant virus was obtained by homologous recombination. In the BmN cells infected with one of the several recombinant viruses, expression of a protein was detected by the SDS-PAGE and Western blotting using an anti-E protein monoclonal as well as anti-JE virus polyclonal antibodies. The molecular weight of the expressed protein was estimated as 53 K daltons, similar to the value of JE virus E protein. Five out of the 9 mice immunized with the recombinant viurs-infected BmN cell homogenate showed weak neutralization activity to JE virus in their sera. In contrast, none of the sera from mice immunized with uninfected cell homogenate showed positive neutralization. The result seems to be the promising first step toward the second generation JE vaccine in the future.
URI: http://hdl.handle.net/10069/4504
ISSN: 03855643
Type: Departmental Bulletin Paper
Appears in Collections:Volume 29, No. 4

Citable URI : http://hdl.handle.net/10069/4504

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