Fujita H, Sumiyoshi H, Mori C, Manabe S, Takagi M, Yoshida I, Morita K, Fuke I, Fukai K, Igarashi A
Bull World Health Organ. 1987;65(3):303-8.
A safe, effective and economical vaccine is required for the prevention of Japanese encephalitis (JE), a disease with high mortality and grave sequelae, which is prevalent in Japan and other countries in east, south-east and southern Asia. As the initial step to produce a second-generation vaccine, recombinant DNA technology was utilized to express the JE virus envelope glycoprotein V3 (E) gene in yeast cells.This report describes the construction of a yeast expression vector in which a cDNA clone covering the V3 gene was connected to the acid-phosphatase promoter of a yeast vector plasmid. Successful expression of the V3 gene was detected by ELISA and Western blotting using monoclonal antibodies against JE V3. Immunization of mice with the V3 antigen expressed in yeast produced limited but definite levels of anti-JE antibodies which could neutralize JE virus. The results are an encouraging step in the development of a practical second-generation JE vaccine.
预防日本脑炎(JE)需要一种安全、有效且经济的疫苗。日本脑炎是一种死亡率高且有严重后遗症的疾病,在日本以及东亚、东南亚和南亚的其他国家流行。作为生产第二代疫苗的第一步,利用重组DNA技术在酵母细胞中表达日本脑炎病毒包膜糖蛋白V3(E)基因。本报告描述了一种酵母表达载体的构建,其中覆盖V3基因的cDNA克隆与酵母载体质粒的酸性磷酸酶启动子相连。使用抗JE V3的单克隆抗体通过ELISA和Western印迹检测到V3基因的成功表达。用酵母中表达的V3抗原免疫小鼠产生了有限但确定水平的抗JE抗体,这些抗体可以中和JE病毒。这些结果是开发实用的第二代JE疫苗中令人鼓舞的一步。