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US 5,175,099 AGranted
Retrovirus-mediated secretion of recombinant products
AssigneeResearch Corporation Technologies, Inc.
InventorJohn W. Wills
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AssigneeResearch Corporation Technologies, Inc.
InventorJohn W. Wills
Estimating the term…
The present invention is directed to replicable expression vectors for producing fusion proteins which are secreted in membraneous particles budded from the cell membrane. In particular these vectors express a hybrid gene product composed of a modified retrovirus gag gene fused to a heterologous gene, or any part thereof, wherein the gag gene modification is sufficient to enable a cell to produce the hybrid gene product in a membraneous particle by budding from the cell membrane into the culture medium or extracellular space, a process known as retrovirus-mediated secretion. The minimum gag sequences needed to obtain particle formation are described. The invention also provides hosts containing the expression vectors, and the fusion proteins produced by the vectors. Further the invention provides the membraneous particles produced by retrovirus-mediated secretion and uses of these particles for protein purification and in therapeutics.
A nucleic acid encoding a hybrid gene comprising at least part of a retrovirus gag gene fused with at least part of a heterologous gene, wherein said gag gene encodes a myristic acid addition site, and wherein said part of said gag gene enables a cell to produce a hybrid gene product in a membraneous particle.
A nucleic acid comprising at least part of an avian retrovirus gag gene that has been modified to encode a myristic acid addition site wherein said gag gene enables a mammalian or an avian cell to produce a gene product in a membraneous particle.
A nucleic acid of claim 18 wherein said avian retrovirus is RSV.
A replicable expression vector comprising p.DELTA.SV.Myr.sub.1, p.DELTA.SV.Myr.sub.1A, pSV.Myr1.R-3K, pSV.Myr1.R-3A, pSV.Myr1.R-3C, pSV.Myr1.R-3J, pSV.Myr1.MA1, pSV.Myr1.Es-Bg, pSV.Myr.3h, pSV.Myr1.PR-A1*, pSV.Myr1.Sm-Bs, pSV.Myr1.DM1, pSV.Myr1.DM2, pSV.Myr1.TM, p.DELTA.SV.Myr.sub.2, pDo.Myr1, pSV.MyCYE, or pSV.MyCYC1.
A method of producing a fusion protein or a fragment thereof comprising:
A method of producing membraneous particles containing a fusion protein or a fragment thereof which comprises:
Dependencies are inferred from the claim text.