High-level expression of the Japanese encephalitis virus E protein by recombinant vaccinia virus and enhancement of its extracellular release by the NS3 gene product.
Recombinant vaccinia viruses expressing the prM and E genes of the Japanese encephalitis virus (JEV) were constructed by use of synthetic promoters. While the recombinant virus mOJ6-SL, with an optimized vaccinia late-gene promoter, produced a 20-fold elevated level of E protein, as well as an 86-kDa precursor protein in infected cells, no significant quantitative difference was detected between the extracellular or cell-surface E protein produced by mOJ6-SL and those produced by mOJ6 with the 7.5-kDa promoter. However, when the cells were infected with Dengue 2 virus before infection with mOJ6-SL, the amount of the extracellular E protein increased 16-fold. In addition, enhancement of its extracellular release was observed when cells were co-infected with mOJ6-SL and recombinant vaccinia virus expressing the NS3 gene of JEV.
Sato
T
T
Biological Science Laboratory, Nippon Zeon Company, Limited, Kanagawa, Japan.
Takamura
C
C
Yasuda
A
A
Miyamoto
M
M
Kamogawa
K
K
Yasui
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003062:Codon / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004664:Encephalitis Virus, Japanese / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D014616:Vaccinia virus / Q000235:genetics*; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
D003062:Codon; D014764:Viral Proteins