Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters fat – fatty oil – ester-type wax – or...
Patent
1996-01-25
1998-06-30
LeGuyader, John L.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters fat, fatty oil, ester-type wax, or...
4352523, 43525411, 514 12, 530324, 530370, 536 236, A01H 500, A01N 6500, C07H 2100, C07K 14415
Patent
active
057736944
DESCRIPTION:
BRIEF SUMMARY
This application claims benefit of international application PCT/GB94/01636, filed Jul. 29, 1994.
This invention relates to antimicrobial proteins, processes for their manufacture and use, and DNA sequences coding for them.
In this context, antimicrobial proteins are defined as proteins possessing at least one of the following activities: antifungal activity (which may include anti-yeast activity); antibacterial activity. Activity includes a range of antagonistic effects such as partial inhibition or death. Such proteins may be oligomeric or may be single peptide subunits.
Various proteins with antimicrobial activity have been isolated from plant sources, and such proteins are often believed to take part in host defence mechanisms directed against invading or competing micro-organisms. Some of the proteins are well-characterised, and their amino acid sequence may be known. In some cases, the cDNA or gene encoding the protein has also been isolated and sequenced.
To keep out potential invaders, plants produce a wide array of antifungal compounds, either in a constitutive or an inducible manner. Several classes of proteins with antifungal properties have now been identified, including: Van Parijs J et al, 1991, Planta, 183, 258-264); Gen Microbiol, 136, 2150-2155; Vigers AJ et al, 1991, Molec Plant-Microbe Interact, 4, 315-323; Woloshuk CP et al, 1991, Plant Cell, 3, 619-628); 42:227-240); Biosci Rep, 6, 19-29; Leah et al, 1991, J Biol Chem, 266, 1564-1573; Carrasco et al, 1981, Eur J Biochem, 116, 185-189; Vernon et al, 1985, Arch Biochem Biophys, 238, 18-29; Stirpe and Barbieri, 1986, FEBS Lett, 195, 1.about.8). be used as biocontrol agents.
Other groups of antimicrobial proteins with activity against plant pathogenic fungi (and often some antibacterial activity) are capable of isolation from certain plant species. We have previously described the structural and antifungal properties of several such proteins, including:
the small-sized cysteine-rich proteins Mj-AMP1 (antimicrobial protein 1) and Mj-AMP2 occurring in seeds of Mirabilis jalapa (Cammue BPA et al, 1992, J Biol Chem, 267:2228-2233; International Application Publication Number WO92/15691 published on 17 Sep. 1992);
Ac-AMP1 and Ac-AMP2 from Amaranthus caudatus seeds (Broekaert WF et al, 1992, Biochemistry, 37:4308-4314; International Application Publication Number WO92/21699 published on 10 Dec. 1992);
Ca-AMP1 from Capsicum annuum, Bm-AMP1 from Briza maxima and related proteins found in other plants including Delphinium, Catapodium, Baptisia and Microsensis species (International Patent Application Publication Number WO94/11511, published 26 May 1994);
Rs-AFP1 (antifungal protein 1) and Rs-AFP2 from seeds of Raphanus sativus (Terras FRG et al, 1992, J Biol Chem, 267:15301-13309) and related proteins such as Bn-AFP1 and Bn-AFP2 from Brassica napus, Br-AFP1 and Br-AFP2 from Brassica rapa, Sa-AFP1 and Sa-AFP2 from Sinapis alba, At-AFP1 from Arabidopsis thaliana, Dm-AMP1 and Dm-AMP2 from Dahlia merckii, Cb-AMP1 and Cb-AMP2 from Cnicus benedictus, Lc-AFP from Lathyrus cicera, Ct-AMP1 and Ct-AMP2 from Clitoria ternatea (International Patent Application Publication Number WO93/05153 published 18 Mar. 1993).
These and other plant-derived antimicrobial proteins are useful as fungicides or antibiotics to improve the disease-resistance or disease-tolerance of crops either during the life of the plant or for post-harvest crop protection. The proteins may be extracted from plant tissue or produced by expression within micro-organisms or synthesised. Exposure of a plant pathogen to an antimicrobial protein may be achieved by application of the protein to plant parts using standard agricultural techniques (eg surface spraying). The proteins may also be used to combat fungal or bacterial disease by expression within plant bodies (rather than just at the surface). The antimicrobial protein may be expressed in an endophyte introduced into plant tissue. DNA encoding the antimicrobial proteins (which may be a cDNA clone, a genomic DNA clone or DNA manufactured using
Broekaert Willem Frans
Cammue Bruno Philippe Angelo
Rees Sarah Bronwen
Larson Thomas G.
LeGuyader John L.
Zeneca Limited
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