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1. (WO2005067478) PRODUCTION DE PARTICULES DU TYPE VIRUS ICOSAEDRIQUE RECOMBINE CHEZ DES PSEUDOMONADES
Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

CLAIMS

1) A Pseudomonad cell that comprises a first nucleic acid construct comprising:
a) at least one nucleic acid sequence encoding a icosahedral viral capsid; and b) at least one nucleic acid sequence encoding a recombinant peptide.
2) The cell of claim 1, wherein the Pseudomonad is Pseudomonas fluorescens.

3) The cell of claim 1, wherein the icosahedral viral capsid is from a virus that does not display a native tropism to a Pseudomonad cell.
4) The cell of claim 3, wherein the icosahedral viral capsid is from a plant
icosahedral virus.
5) The cell of claim 4, wherein the plant icosahedral virus is selected from the group consisting of a Cowpea Chlorotic Mottle Nirus, a Cowpea Mosaic Nirus, and an Alfalfa Mosaic Nirus.
6) The cell of claim 1, wherein the nucleic acid encodes at least two different icosahedral viral capsids.
7) The cell of claim 6, wherein at least one of the icosahedral viral capsids is
from a plant icosahedral virus.
8) The cell of claim 1, wherein the nucleic acid encoding the recombinant
peptide contains more than one monomer.
9) The cell of claim 8, wherein the nucleic acid encoding the recombinant
peptide contains at least three monomers.
10) The cell of claim 8, wherein the monomers are operably linked as concatamers.

11) The cell of claim 1 , wherein the recombinant peptide fused to the icosahedral capsid is a therapeutic peptide.
12) The cell of claim 1, wherein the recombinant peptide is an antigen.
13) The cell of claim 12, wherein the antigen is selected from the group consisting of a Canine Parvovirus antigen, a Bacillus Anthracis antigen, and an Eastern
Equine Encephalitis viral antigen.
14) The cell of claim 1 , wherein the recombinant peptide is an antimicrobial
peptide.
15) The cell of claim 14, wherein the antimicrobial peptide is selected from the group consisting of D2A21 and PBF20.

16) The cell of claim 1, wherein the recombinant peptide is at least 7 amino acids in length.
17) The cell of claim 16, wherein the recombinant peptide is at least 15 amino
acids in length.
18) The cell of claim 1, wherein the cell further comprises a second nucleic acid encoding a wild type icosahedral viral protein.
19) The cell of claim 1, wherein the cell further comprises a second nucleic acid comprising:
c) at least one nucleic acid sequence encoding a second icosahedral viral
capsid; and
d) at least one nucleic acid sequence encoding a second recombinant peptide.

20) The cell of claim 19 wherein the first and second icosahedral viral capsids are different.
21) A Pseudomonad cell that comprises a fusion peptide, wherein the fusion
peptide comprises:
a) at least one icosahedral viral capsid; and
b) at least one recombinant peptide.
22) The cell of claim 21 wherein the fusion peptide assembles within the cell to form a virus like particle.
23) The cell of claim 21 wherein the fusion peptide assembles within the cell to form a soluble cage structure.
24) The cell of claim 22, wherein the virus like particle is not capable of
replication.
25) The cell of claim 22, wherein the virus like particle is not capable of infecting a cell.
26) The cell of claim 21, wherein the recombinant peptide is inserted into at least one surface loop of the icosahedral capsid.
27) The cell of claim 21 , wherein a recombinant peptide is inserted into more than one surface loop of the icosahedral capsid.
28) The cell of claim 21 , wherein the fusion peptide comprises more than one
recombinant peptide fused to an icosahedral viral capsid.
29) The cell of claim 28, wherein the recombinant peptides are different.
30) The cell of claim 21, wherein the recombinant peptide is a therapeutic peptide.

31) The cell of claim 21 , wherein the recombinant peptide is an antigen.

32) The cell of claim 31 , wherein the antigen is selected from the group consisting of a Canine Parvovirus antigen, a Bacillus Anthracis antigen, and an Eastern
Equine Encephalitis viral antigen.
33) The cell of claim 22, wherein the virus like particle is capable of use as a
vaccine.
34) The cell of claim 21 , wherein the recombinant peptide is a peptide that is an antimicrobial peptide.
35) The cell of claim 34, wherein the antimicrobial peptide is selected from the group consisting of D2A21 and PBF20.
36) The cell of claim 21 , wherein the recombinant peptide is at least 7 amino acids in length.
37) The cell of claim 21, wherein the recombinant peptide is at least 15 amino acids in length.
38) The cell of claim 21 , wherein the cell further comprises a wild type
icosahedral viral capsid.
39) The cell of claim 21 , wherein the cell further comprises a second fusion
peptide comprising:
a) at least a second icosahedral viral capsid; and
b) at least a second recombinant peptide.
40) The cell of claim 39 wherein the second fusion peptide assembles within the cell to form a virus like particle or a soluble cage structure.
41) The cell of claim 39 wherein the second fusion peptide comprises a different amino acid sequence than the first fusion peptide.
42) The cell of claim 21 wherein the viral capsid and the recombinant peptide are linked by an amino acid sequence comprising a linker.
43) The cell of claim 42 wherein the linker amino acid sequence comprises a cleavable sequence.
44) The cell of claim 21 , wherein the Pseudomonad is Pseudomonas fluorescens.

45) A nucleic acid construct comprising a first nucleic acid sequence encoding an icosahedral viral capsid operably linked to a second nucleic acid sequence encoding a peptide that is toxic to a microbial cell.
46) The construct of claim 45, wherein the icosahedral viral capsid is from a plant icosahedral virus.

47) The construct of claim 46, wherein the plant icosahedral virus is selected from the group consisting of a Cowpea Chlorotic Mottle Nirus, a Cowpea Mosaic
Nirus, and an Alfalfa Mosaic Nirus.
48) The construct of claim 46, wherein the toxic peptide comprises more than one peptide monomer sequence.
49) The construct of claim 46, wherein the toxic peptide comprises at least three peptide monomer sequences.
50) The construct of claim 48, wherein the monomers are operably linked to form a concatamer.
51) The construct of claim 45, wherein the operable linkage is internal to the first nucleic acid sequence encoding the capsid.
52) The construct of claim 45 wherein the second nucleic acid sequence encoding the toxic peptide is operably linked to the capsid sequence in a location
encoding for at least one surface loop of the capsid.
53) The construct of claim 45, wherein the construct encodes more than one toxic peptide sequence operably linked to the capsid sequence locations encoding for more than one surface loop of the capsid.
54) The construct of claim 45, wherein the recombinant peptide is an antimicrobial peptide.
55) The construct of claim 54, wherein the antimicrobial peptide is selected from the group consisting of D2A21 and PBF20.
56) A process for producing a recombinant peptide comprising:
a) providing a Pseudomonad cell;
b) providing a nucleic acid encoding a fusion peptide, wherein the fusion
peptide comprises at least one recombinant peptide and at least one
icosahedral capsid;
c) expressing the nucleic acid in the Pseudomonad cell, wherein the fusion
peptide assembles into virus like particles; and
d) isolating the virus like particles.
57) The process of claim 56, further comprising:
e) cleaving the fusion peptide to separate the recombinant peptide from the icosahedral viral capsid.
58) The process of claim 56, wherein the Pseudomonad is Pseudomonas
fluorescens.

59) The process of claim 56, wherein the virus like particle is not capable of replication.
60) The process of claim 56, wherein the virus like particle is not capable of infecting a cell.
61) The process of claim 56, wherein the icosahedral viral capsid is from a virus that does not display a native tropism to a Pseudomonad cell.
62) The process of claim 56, wherein the icosahedral viral capsid is from a plant icosahedral virus.
63) The process of claim 62, wherein the plant icosahedral virus is selected from the group consisting of a Cowpea Chlorotic Mottle Nirus, a Cowpea Mosaic Nirus, and an Alfalfa Mosaic Nirus.
64) The process of claim 56, wherein the nucleic acid comprises a nucleic acid sequence encoding at least two different icosahedral viral capsids.
65) The process of claim 64, wherein at least one of the icosahedral viral capsids is from a plant icosahedral virus.
66) The process of claim 56, wherein the recombinant peptide comprises more than one peptide monomer.
67) The process of claim 56, wherein the recombinant peptide comprises at least three monomers.
68) The process of claim 66, wherein the monomers are operably linked as a concatamer.
69) The process of claim 56, wherein the recombinant peptide is operably linked to at least one surface loop of the icosahedral capsid.
70) The process of claim 69, wherein a recombinant peptide is operably linked to more than one surface loop of the icosahedral capsid.
71) The process of claim 56, wherein the fusion peptide comprises more than one recombinant peptide, the recombinant peptides being dissimilar.
72) The process of claim 56, wherein the recombinant peptide is a therapeutic peptide.
73) The process of claim 56, wherein the recombinant peptide is an antigen.

74) The process of claim 73, wherein the antigen is selected from the group
consisting of a Canine Parvovirus antigen, a Bacillus Anthracis antigen, and an Eastern Equine Encephalitis viral antigen.

75) The process of claim 56, wherein the virus like particle is capable of use as a vaccine.
76) The process of claim 56, wherein the recombinant peptide is a peptide that is an antimicrobial peptide.
77) The process of claim 76, wherein the antimicrobial peptide is selected from the group consisting of D2A21 and PBF20.
78) The process of claim 56, wherein the recombinant peptide is at least 7 amino acids in length.
79) The process of claim 56, wherein the recombinant peptide is at least 15 amino acids in length.
80) The process of claim 56, wherein the cell further comprises a second nucleic acid encoding a wild type icosahedral viral capsid.
81) The process of claim 56, wherein the cell further comprises a second nucleic acid encoding a second fusion peptide comprising:
a) at least a second icosahedral viral capsid; and
b) at least a second recombinant peptide.
82) The process of claim 81 comprising expressing the second nucleic acid in the cell.
83) The process of claim 81 wherein the second fusion peptide assembles within the cell to form a virus like particle or a soluble cage structure.
84) The process of claim 81 wherein the first icosahedral viral capsid comprises a first amino acid sequence and the second icosahedral viral capsid comprises a second amino acid sequence and the first and second capsid sequences are different.
85) The process of claim 81 wherein the first recombinant peptide comprises a first amino acid sequence and the second recombinant peptide comprises a second amino acid sequence and the first and second recombinant peptide sequences are different.