Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part
Patent
1995-02-02
1999-10-05
Lau, Kawai
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
Higher plant, seedling, plant seed, or plant part
435 6, 435 691, 4352523, 43525411, 4353201, 424 932, 424 935, 536 232, C12N 514, C12N 115, C12N 121, A01N 6304
Patent
active
059627654
ABSTRACT:
We have studied the regulation of the extracellular chymoelastase protease (Pr1) of Metarhizium anisopliae, an enzyme involved in the penetration of insect cuticle by Metarhizium and other entomopathogenic fungi. We report here the isolation and characterization of a Pr1 cDNA clone with a full length insert. Pr1 is synthesized as a large precursor (40.3 kDa) containing a signal peptide and a propeptide and the mature protein is predicted to have a relative molecular mass of 28.6 kDa. The primary structure of Pr1 shares extensive homology (30-60%) with enzymes of the subtilisin subclass of the serine endopeptidases and the serine, histidine and aspartyl components of the active site in subtilisins are preserved. The genes coding for chymoelastase or slightly altered versions thereof, can be used to transform various organisms (i.e. fungi, viruses, plants, bacteria, etc.) such that the transformed organisms are capable of producing chymoelastase in recoverable quantities. Fragments and derviatives of a DNA sequence coding for a chymoelastase could be used to code for a polypeptide having an activity which can: a) bind to insect cuticle; b) enhance signal processing of proteins; c) hydrolyse polypepetides; d) suppress protease expression; or e) be used as a probe to identify homologous genes in organisms. While chymoelastases and Pr1 have been previously isolated, new and novel uses for chymoelastase are disclosed, wherein the chymoelastase is used to selectively degrade protein in the presence of non-protein polymers. A new insecticide insecticide is disclosed which comprises a recombinant virus, microorganism, cell, plant or fungi infects, is eaten by or otherwise taken up by, an insect and expresses the enzyme Pr1 within said insect such that Pr1 activates a prophenoloxidase system within said insect.
REFERENCES:
patent: 4987077 (1991-01-01), Charnley et al.
patent: 5266317 (1993-11-01), Tomalski et al.
Gupta, A.S. et al, 1993, Increased Resistanct To Oxidative Stress In Transgenic Plants That Overexpress chloroplastic Cu/Zu Superoxide dismutase, Proc. Natl. Acad. Sci, USA, vol. 90, pp. 1629-1633.
Clare, B.G. et al, 1990, Agrobacterium in Plant Disease, Biological Disease Control and Plant Genetic Engineering, Sci. Progress, 74: 1-13.
Binns, A.N. et al, 1988, Cell Biology of Agrobacterium Infection and Transformation of Plants, Ann. Rev. Microbiol., 42: 575-606.
Betzel, C. et al, Active-site geometry of proteinase K, FEBS Letters vol. 197, pp. 105-114, (1986).
Carter, P. et al, Dissecting the catalytic triad of a serine protease, Nature, vol. 332 (1988) pp. 564-568.
Charnley, A.K. et al, The Role of Cuticle-Degrading Enzymes in Fungal Pathogenesis in Insects, Plenum Press, pp. 267-286, (1991).
Davidow, L.S. et al, Cloning and Sequencing of the Alkaline Etracellular Protease Gene of Yarrowia lipolytica, Jnl. of Bact., (Oct. 1987), pp. 4621-4629.
Deane, S. et al, Nucleotide sequence of the Vibrio alginolytieus calcium-dependent, detergent-resistant alkaline serine exoprotease A, Gene, 76 (1989) pp. 281-288.
Egeling, W. et al, Proteinase K from Tritirachium album Limber, Eur.J. Biochem, 47, (1974) pp. 91-97.
Flurkey, W.H. et al, In Vitro Translation of Cutinase mRNA: Evidence for a Precursor Form of an Extracellular Fungal Enzyme, Archi. of Bioch and Biophysis, (1981) vol. 2112, pp. 154-161.
Goettel, M.S. et al, Ultrastructural Localization of a Cuticle-degrading Protease Produced by the Entromopathogenic Fungus Metarhizium anisopliae during Penetration of Host (Manduca sexta) Cuticle, Jnl of Gen Microb. (1989) 135, 2233-2239.
Goettel, M.S. et al., Pathogenicity and growth of Metarhizium anisopliae stably transformed by benomyl resistance, Curr Genet (1990) 17: 129-132.
Gunkel, F.A. et al, Proteinase K from Tritirachium album Limber, Eur. J. Biochem. (1989) 179, 185-194.
Heale, J.B. et al, Prospects for Strain Improvement in Entomopathogenic Fungi, Pestic. Sci. (1989) 26: 79-92.
Heale, J.B., The Potential Impact of fungal genetics and molecular biology on biological control, with particular reference to entomopathogens, pp. 2121-234, (1991).
Jany, K. et al, Amino acid sequence of proteinase K from the mond Tritirachium album Limer, Elsevier Sci. Publ., (Apr. 1986) vol. 199:139-144.
Kaneko, R. et al, Molecular Cloning of athe Structural Gene for Alkaline Elastase YaB, a New Subtilisin Produced by an Alkalophilic Bacillus Strain, Jnl of Bacter., (Sep. 1989) pp. 5232-5236.
Hepburn, H.F., Structure of the Integument, Pergamon Press, vol. 3, pp. 1-58, (1991).
Kessler, E. et al, Synthesis, Processing, and Transport of Pseudomonas aeruginosa Elastase, Jnl. of Bacter., (Nov. 1988) pp. 5241-5247.
Gurr, S.J. et al, The structure and organization of nuclear genes of filamentous fungi, Soc. Gen Microbiology, vol. 22, (1987) pp. 93-139.
Kramer, K.J. et al, Insect Cuticle Structure and Metabolism, American Chemical Soc., (1987) Chaper 12, pp. 160-185.
Kraut, J., Serine Proteases: Structure and Mechanism of Catalysis, Ann. Rev. Biochem. (1977) 46:331-358.
Laemmli, U.K., Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4, Nature, vol. 227, (1970) pp. 680-685.
DeLorenzo, G. et al, Induction of Extracellular Polygalacturonase and Its mRNA in the Phytopathogenic Fungus Fusarium moniliforme, Jnl. Gen. Microbiology, (1987) 133: 3365-3373.
Lovett, J.S., The Molocular Biology of Fungal Development, vol. 2, Macmillar Publ. Co., pp, 64-100, (1991).
Maniatis, T. et al, Molecular Cloning-A Laboratory Manual, (1982), Cold Spring Harbor Laboratory, pp. 194-195.
Matsuzawa, H. et al, Purification and characterization of aqualysin I (a thermophilic alkaline serine protease) produced by Thermus aquatics YT-1, Eur. J. Biochem. (1988) 171: 441-447.
McCoy, C.W. et al, Dntomogenous Fungi, vol V., Part A., CRC Press, Inc., pp. 151-236, (1991).
Moehle, C.M. et al, Protease B. of the Tysosomelike Vacuole of the Yeast Sccharomyces cerevisiae is homologous to the Subtilisin Family of Serine Proteases, Mole and Cell Biol., (Dec. 1987) pp. 4390-4399.
Moehle, C.M. et al, Processing Pathway for Protease B. of Saccharomyces cerevisiae, Jnl of Cell Biol., vol 108, (Feb. 1989) pp. 309-324.
Roberts, D.W. et al, Use of Pathogens in Insect Control, vol. II, CRC Press, Inc., pp. 243-278, (1991).
Roberts, D.W., World Picture of Biological Control of Isnects by Fungi, Mem. Inst. Oswaldo Cruz, Rio de Janeiro, Bol. 85, (1989) pp. 89-99.
Robets, D.W et al, Entomogenous Fungi, Biol. of Conidial Fung, vol. 2, (1981), pp. 201-235.
Sambrook, J. et al, Identification of cDNA Clones of Interest, Cold Spring Harbor Lab. Press, pp. 8.46-8.86, (1991).
Samal, B.B. et al, Cloning and expression of the gene encoding a novel proteinase from Tritirachium album Limer, (1989), Gene. 85:329-333.
Sive, H.L. et al, A simple subtractive hybridization technique employing photoactivatable biotin and phenol extraction, Nucl Acid Res., vol.16 (1988).
Tatsumi, H. et al, A full length cDNA clone for th alkaline protease from Aspergillus oryzae: Strctural analysis and expression in Saccharomyces cerevisiae, Mol. Gen. Genet (1989) 219:33-38.
St.Leger, R.J., (1990), The role of cuticle-degrading enzymens in fungal pathogenesis of insects. In Vth Intern. Colloquium on Invertebrate Pathology and Microbial Control, Preceedings held Adelaide, Australia, pp. 308-312.
St.Leger, R.J. et al, (1991), Kinetics of the digestion of insect cuticles by a protease (Pr1) from Metarhizium anisopliae, J. Invertebr. Pathol. 57: 146-147.
St.Leger, R.J. et al, (1991), Characterization of chitinolytic enzymes produced by the entomopathogen Metarhizium anisopliae., J. Invertebr. Pathol. 58: 415-426.
St.Leger, R.J. et al, (1991), Prepenetration events during infection of host cuticle by Metarhizium anisopliae, J. Invert. Pathol., 58:168-179.
St.Leger, R.J. et al, (1992), Fungi for the biocontrol of insects: Tools and trends. In Molecular Biol. of Filamentous Fungs, pp. 45-63.
Staples, R.C. et al, (1989), Strategies for genetic engineering of fungal entomopathogens. In Biothechnology, Biological Pesticides and Novel Plant-Pest Resistance for Insect Pest Ma
Roberts Donald W.
St.Leger Raymond J.
Staples Richard C.
Boyce Thompson Institute for Plant Research Inc.
Lau Kawai
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