US2001014732A1PendingUtilityA1
Biocidal proteins
Est. expiryAug 29, 2011(expired)· nominal 20-yr term from priority
Inventors:Willem BroekaertBruno P. A. CammueRupert William OsbornSarah Bronwen ReesFranky TerrasJozef Vanderleyden
A01N 65/08C12N 15/8281A01N 37/46C07K 14/415C12N 15/8282A01N 65/12
51
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Claims
Abstract
Biocidal proteins isolated from seeds have been characterized, in particular proteins isolated from members of the Brassicaceae, Compositae and Leguminosae families including Raphanus, Brassica, Sinapis, Arabidopsis, Dahlia, Cnicus, Lathyrus and Clitoria. The proteins show a wide range of antifungal activity and some are active against Gram-positive bacteria. All share a common amino acid sequence. DNA encoding the proteins has been isolated and incorporated into vectors. Plants transformed with this DNA may be produced. The proteins find commercial application as antifungal or antibacterial agents; transformed plants will show increased disease-resistance.
Claims
exact text as granted — not AI-modified1 . An antimicrobial protein having substantially the amino acid sequence shown in FIGS. 27 to 29 or in FIG. 32.
2 . A protein as claimed in claim 1 which is an oligomer and which comprises at least one polypeptide having substantially the amino acid sequence shown in FIGS. 27 to 29 or in FIG. 32.
3 . A protein as claimed in claim 1 or claim 2 which is capable of being isolated from a plant seed.
4 . A protein as claimed in claim 3 which is capable of being isolated from a seed of the family Brassicaceae or of the family Compositae or of the family Leguminosae.
5 . A protein as claimed in claim 4 which is isolated from Raphanus, Brassica, Sinapis, Arabidopsis, Dahlia, Cnicus, Lathyrus or Clitoria.
6 . A pure protein Rs-AFP1, capable of being isolated from Raphanus seed.
7 . A pure protein Rs-AFP2, capable of being isolated from Raphanus seed.
8 . A pure protein Rs-nsLTP, capable of being isolated from Raphanus seed.
9 . Pure proteins Bn-AFP1, Bn-AFP2, Br-AFP1 and Br-AFP2, capable of being isolated from Brassica seed.
10 . Pure proteins Sa-AFP1 and Sa-AFP2, capable of being isolated from Sinapis seed.
11 . A pure protein At-AFP1, capable of being isolated from Arabidopsis seed.
12 . A pure protein Dm-AMP1, capable of being isolated from Dahlia seed.
13 . A pure protein Dm-AMP2, capable of being isolated from Dahlia seed.
14 . A pure protein Cb-AMP1, capable of being isolated from Cnicus seed.
15 . A pure protein Cb-AMP2, capable of being isolated from Cnicus seed.
16 . A pure protein Lc-AFP, capable of being isolated from Lathyrus seed.
17 . A pure protein Ct-AMP1, capable of being isolated from Clitoria seed.
18 . A pure protein Ct-AMP2, capable of being isolated from Clitoria seed.
19 . A protein as claimed in any of claims 1 to 18 which is synthetic.
20 . A recombinant DNA sequence encoding a protein as claimed in any of claims 1 - 19 .
21 . A DNA sequence as claimed in claim 20 which is a cDNA.
22 . A DNA sequence as claimed in claim 20 which is genomic DNA.
23 . A DNA sequence as claimed in claim 22 which is isolated from a plant genome.
24 . A DNA sequence as claimed in claim 23 which includes a promoter sequence.
25 . A promoter sequence obtainable from a gene encoding a protein as claimed in any of claims 1 - 19 .
26 . A vector containing a DNA sequence as claimed in claim 20 .
27 . A biological system including recombinant DNA as claimed in claim 20 such that the encoded protein is expressed.
28 . A biological system as claimed in claim 27 which is a micro-organism.
29 . A biological system as claimed in claim 27 which is a plant.
30 . An antimicrobial protein produced by expression of recombinant DNA as claimed in claim 20 .
31 . A plant transformed with recombinant DNA as claimed in claim 20 .
32 . A plant as claimed in claim 26 in which the recombinant DNA encodes at least one of the following proteins: Rs-AFP1, Rs-AFP2, Rs-nsLTP, Bn-AFP1, Bn-AFP2, Br-AFP1, Br-AFP2, Sa-AFP1, Sa-AFP2, At-AFP1, Dm-AMP1, Dm-AMP2, Cb-AMP1, Cb-AMP2, Lc-AFP, Ct-AMP1, Ct-AMP2.
33 . Seeds and progeny of a plant as claimed in claim 31 or claim 32 .
34 . A composition containing at least one of the proteins as claimed in any of claims 1 to 19 or claim 30 .
35 . A process of combating fungi or bacteria which comprises exposure to a protein or composition as claimed in any of claims 1 to 19 , claim 30 or claim 34 .
36 . A process of combating fungi or bacteria which comprises exposure to a protein encoded by pea gene pI39, by pea gene pI230, by cowpea gene pSAS10, or by potato gene pI322.
37 . A process of combating fungi or bacteria which comprises exposure to SIα2, γ-1-purothionin, or another α-amylase inhibitor protein.
38 . An extraction process for producing a protein as claimed in any of claims 1 to 19 or claim 30 from organic material containing them which comprises submitting the organic material to maceration and solvent extraction.
39 . An extraction process as claimed in claim 38 where the protein is subsequently purified by centrifugation, chromatography and dialysis.
40 . An extraction process as claimed in either claim 38 or claim 39 where the organic matter comprises seeds of Raphanus, Brassica, Sinapis, Arabidopsis, Dahlia, Cnicus, Lathyrus or Clitoria.
41 . An extraction process as claimed in either claim 38 or claim 39 where the organic matter comprises a biological system as claimed in claim 27 .
42 . A process for producing a protein as claimed in any of claims 1 to 19 which comprises chemical synthesis of the protein.
43 . A process for producing a protein as claimed in any of claims 1 to 19 which comprises expression of a recombinant DNA sequence encoding the protein.Join the waitlist — get patent alerts
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