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Display of heterologous proteins on gp64null baculovirus virions and enhanced budding mediated by a VSV G-stem construct.

Zhou J, Blissard GW

Boyce Thompson Institute, Cornell University, Ithaca, New York 14853; Department of Entomology, Cornell University, Ithaca, NY 14853.

The Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) GP64 envelope glycoprotein is essential for virus entry and plays an important role in virion budding. An AcMNPV construct that contains a deletion of the gp64 gene is unable to propagate infection from cell to cell and this defect results from both a severe reduction in the production of budded virions (BV), and the absence of GP64 on virions. In the current study, we examined GP64 proteins containing N- and C-terminal truncations of the ectodomain and identified a minimal construct capable of targeting the truncated GP64 to budded virions. The minimal budding and targeting construct of GP64 contained 38 amino acids from the mature N terminus of the GP64 ectodomain and 52 amino acids from the C terminus of GP64. Because the Vesicular Stomatitis Virus (VSV) G protein was previously found to rescue infectivity of a gp64null AcMNPV virus, we also examined a small C-terminal construct of the VSV G protein. We found that a construct containing 91 amino acids from the C terminus of VSV G (termed G-stem) was capable of rescuing AcMNPV gp64null virion budding to wild type (wt) or near wt levels. We also examined display of chimeric proteins on the gp64null AcMNPV virion. By generating viruses that expressed chimeric influenza hemagglutinin (HA) proteins containing the GP64 targeting domain, and co-infecting those viruses with a virus expressing the G-stem construct, we demonstrate enhanced display of the HA protein on gp64null AcMNPV budded virions. The combined use of gp64null virions, VSV G-stem enhanced budding, and GP64 domains for targeting heterologous proteins to virions should be valuable for biotechnological applications ranging from targeted transduction of mammalian cells to vaccine production.

Published 8 November 2007 in J Virol.
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