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Production of Slowly Sedimenting and Rapidly Sedimenting Components Associated with Japanese Encephalitis Virus Envelope Glycoprotein E in Infected Cell Culture Fluids


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タイトル: Production of Slowly Sedimenting and Rapidly Sedimenting Components Associated with Japanese Encephalitis Virus Envelope Glycoprotein E in Infected Cell Culture Fluids
著者: Shameem, Golam Masud Mohammad / Morita, Kouichi / Bundo-Morita, Keiko / Igarashi, Akira
発行日: 1989年 9月30日
出版者: 長崎大学熱帯医学研究所 / Institute of Tropical Medicine, Nagasaki University
引用: 熱帯医学 Tropical medicine 31(3). p111-123, 1989
抄録: Production of Japanese encephalitis (JE) virus glycoprotein E into culture fluids of infected mosquito (C6/36) and baby hamster kidney (BHK21) cells at 28 and 37℃ was investigated by the sandwich ELISA using an anti-E monoclonal antibody (MCA). Besides rapidly sedimenting component (RE) at the position of complete virion, slowly sedimenting component (SE) was found near the top of the sucrose gradient sedimentation. Except the first 24 hr of infection when virus production was not so much at 28℃, the peak of SE was more pronounced when the infected cells were incubated at 28 than at 37℃. Production of SE was generally more remarkable in C6/36 than in BHK21 cells at both temperatures. At 28℃, production of both SE and RE continued until 72 hr in C6/36 and 60 hr in BHK21 cells. While, production of both SE and RE declined after 36 hr in both cell lines at 37℃. When SE and RE were recentrifuged, they appeared as independent components. Although both SE and RE were accompanied by the peak of hemagglutination activity (HA), the HA of SE was relatively lower than that of RE by comparing the ELISA color development. Western blotting after sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a single band with apparent Mw of the E protein (54K) for both SE and RE fractions. Electron microscopy (EM) with negative staining revealed spherical particles of 45-55 nm in diameter corresponding to the complete virion together with components of 5-10 nm diameter in RE fractions. While in SE fractions, various components of 5-20 nm in diameter, apparently their aggregates, and membranous structures were observed.
URI: http://hdl.handle.net/10069/4549
ISSN: 03855643
資料タイプ: Departmental Bulletin Paper
出現コレクション:第31巻 第3号

引用URI : http://hdl.handle.net/10069/4549

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