(NAS Colloquium) Genetic Engineering of Viruses and Viral Vectors (1996) / Chapter Skim
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Foreign glycoproteins expressed from recombinant vesicular stomatitis viruses are incorporated efficiently into virus particles
Foreign glycoproteins expressed from recombinant vesicular stomatitis viruses are incorporated efficiently into virus particles
Pages 11359-11365
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From page 11359... ...
Incorporation of foreign viral spike glycoproteins into the envelope of recombinant VSV particles might allow development of a new generation of killed virus vaccines or retargeting of virus particles to specific cell types. Also, expression of other membrane proteins from the VSV genome and their subsequent incorporation into the virions could yield large quantities of the expressed protein in highly purified form for structural or functional studies.
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From page 11360... ...
Chloramphenicol acetyl transferase expression employed the VSV vector pVSV-XN1 in which a new transcription unit and cloning site were inserted between the G and L genes of VSV (19. In the present study we have used this same vector to derive recombinant VSVs expressing four different membrane proteins that are transported to the plasma membrane and we have examined their incorporation into the membrane of particles budding from the cell surface.
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From page 11361... ...
Although biochemical analysis of purified virions showed incorporation of CD4 and CD4G into VSV, it was unclear if every particle contained the foreign protein or if there were changes to the structure of VSV virions as a result of the inclusion of an additional gene and protein. To address these questions directly, purified wt VSV, VSVCD4, and VSV-CD4G virions were examined by electron microscopy using Immunogold-coupled antibodies and negative stain (Fig.
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From page 11362... ...
Because the cellular CD4 protein was incorporated well into VSV virions, we then wished to determine if glycoproteins from another virus could also be expressed in VSV recombinants and incorporated into the VSV viral envelope. For these experiments we chose measles virus, a negative strand RNA virus from the distantly related paramyxovirus group.
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From page 11363... ...
We have reported here that both viral and nonviral proteins can be incorporated at high levels along with the VSV glycoprotein into the envelope of VSV. This high level incorporation is achieved by expressing the foreign proteins from genes inserted directly into the VSV genome, ~_'~ ~ f':^'=~:> ~ ~ ~.: 11~1 1 : ~d ~ 1 · ~ l :1 1 : -H f'.-G .'N .
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From page 11364... ...
reduce the efficiency of glycoprotein incorporation into the virus particle suggesting an important interaction occurring through the glycoprotein tail, presumably with the matrix protein. The results presented here illustrate that VSV particles can incorporate several unrelated foreign membrane proteins into the viral envelope at levels up to 30% of G protein itself.
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From page 11365... ...
Colloquium Paper: Schnell of a/.
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