Home Acta Virologica 2004 Acta Virologica Vol.48, p.97-107, 2004

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Quarterly,
Founded: 1957
ISSN 0001-723X
E-ISSN 1336-2305

Published in English

Impact Factor = 1.82

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Acta Virologica Vol.48, p.97-107, 2004

Title: EXPRESSION OF HERPES SIMPLEX VIRUS 1 GLYCOPROTEIN D IN PROKARYOTIC AND EUKARYOTIC CELLS
Author: T. MOSKO, J. KOSOVSKÝ, I. REZUCHOVÁ, V. ĎURMANOVÁ, M. KÚDELOVÁ, J. RAJČÁNI
Abstract: Recombinant plasmids encoding either the full-length glycoprotein D (FLgD) or truncated gDs were constructed. The recombinant plasmids were expressed in Escherichia coli and BHK-21 cells. The strongest expression was obtained with the recombinant plasmid encoding a truncated gD which corresponded to the gD ectodomain. The cells transformed with this plasmid showed good exponential growth ensuring satisfactory yields of the expressed polypeptide in the form of the fusion protein. The fusion protein was biotinylated and efficiently purified. The shortest truncated gD, which contained the main continuous antigenic locus VII binding neutralization antibody and additional continuous antibody binding epitopes, still reacted with specific antibody as proven by immunoblot analysis. In addition, a shuttle vector for expression of FLgD in mammalian cells was constructed. This vector-transfected BHK-21 cells expressed gD for 40 days during 9 consecutive passages. The expression of gD began on day 2 and culminated at day 9 post transfection (p.t.).
Keywords: BHK-21 cells; Escherichia coli; expression; fusion protein; glycoprotein D; Herpes simplex virus 1; recombinant plasmid; shuttle vector
Year: 2004, Volume: 48, Issue: Page From: 97, Page To: 107


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