University of Florida Research Foundation Inc

Yi Li

Richard W Moyer

Richard L Hall

Michael E Gruidl

The subject invention pertains to recombinant entomopox vectors which are useful for the delivery and stable expression of heterologous DNA in vertebrate cells. Specifically exemplified is a recombinant EPV from(AmEPV). Because of the capacity of the EPV to incorporate foreign or heterologous DNA sequences, the vectors of the subject invention can be used to deliver DNA inserts that are larger than 10 kb in size. Accordingly, one aspect of the present invention concerns use of the recombinant vectors for delivery and expression of biological useful proteins in gene therapy protocols. In addition, the subject invention concerns novel AmEVP polypeptides and the polynucleotide sequences which encode these polypeptides.

Materials and methods for delivery and expression of heterologous DNA in vertebrate cells

The subject invention pertains to novel Entomopoxvirus (EPV) polynucleotide sequences free from association with other viral sequences with which they are naturally associated, recombinant polynucleotide vectors containing the sequences, recombinant viruses containing the sequences, and host cells infected with the recombinant viruses are provided herein, as well as methods for use thereof in the expression of heterologous proteins in both insect and mammalian host cells.

Entomopoxvirus expression system

The subject invention pertains to recombinant entomopox vectors which are useful for the delivery and stable expression of heterologous DNA in vertebrate cells. Specifically exemplified is a recombinant EPV from amsacta moorei (AmEPV). Because of the capacity of the EPV to incorporate foreign or heterologous DNA sequences, the vectors of the subject invention can be used to deliver DNA inserts that are larger than 10 kb in size. Accordingly, one aspect of the present invention concerns use of the recombinant vectors for delivery and expression of biological useful proteins in gene therapy protocols.

Materials and methods for delivery and expression of heterologous DNA in

This invention provides recombinant entomopoxvirus as a novel vector and method of using such vector for delivery and expression of genes of biological significance to vertebrate cells, including human cells. This invention includes the use of an early entomopoxvirus or like gene promoter to drive the expression of an heterologous gene product in a vertebrate cell infected with the recombinant entomopoxvirus vector. We have discovered that, while the entomopoxvirus vector of this invention does not replicate in the vertebrate cell, it enters vertebrate cell cytoplasm and achieves expression of an heterologous gene inserted into the rEPV without causing cytopathic or cytotoxic effects. We further provide a method for expression of the heterologous gene under control of late, strong entomopoxvirus gene promoters, vertebrate gene or viral promoters, and for stable integration of the heterologous gene delivered by the rEPV into the vertebrate cell.

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Gainesville, FL, United States of America

Yi Li