Chimeric Isoprenoid Synthases and Uses Thereof Joseph Chappell University of Kentucky, [email protected]

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Chimeric Isoprenoid Synthases and Uses Thereof Joseph Chappell University of Kentucky, Chappell@Uky.Edu University of Kentucky UKnowledge Pharmaceutical Sciences Faculty Patents Pharmaceutical Sciences 10-20-1998 Chimeric Isoprenoid Synthases and Uses Thereof Joseph Chappell University of Kentucky, [email protected] Kyoungwhan Back University of Kentucky Right click to open a feedback form in a new tab to let us know how this document benefits oy u. Follow this and additional works at: https://uknowledge.uky.edu/ps_patents Part of the Pharmacy and Pharmaceutical Sciences Commons Recommended Citation Chappell, Joseph and Back, Kyoungwhan, "Chimeric Isoprenoid Synthases and Uses Thereof" (1998). Pharmaceutical Sciences Faculty Patents. 101. https://uknowledge.uky.edu/ps_patents/101 This Patent is brought to you for free and open access by the Pharmaceutical Sciences at UKnowledge. It has been accepted for inclusion in Pharmaceutical Sciences Faculty Patents by an authorized administrator of UKnowledge. For more information, please contact [email protected]. US005 824774A United States Patent [19] [11] Patent Number: 5,824,774 Chappell et al. [45] Date of Patent: Oct. 20, 1998 [54] CHIMERIC ISOPRENOID SYNTHASES AND Chappell et al., “Accumulation of Capsidiol in Tobacco Cell USES THEREOF Cultures Treated With Fungal Elicitor,” Phytochemistry 26:2259—2260 (1987). [75] Inventors: Joseph Chappell; Kyoungwhan Back, Chappell, “Biochemistry and Molecular Biology of the both of Lexington, Ky. Isoprenoid Biosynthetic PathWay in Plants,” Annu. Rev. Plant Physiol. Plant Mol. Biol. 46:521—547 (1995). [73] Assignee: Board of Trustees of the University of Chappell et al., “Is the Reaction Catalyzed by Kentucky, Lexington, Ky. 3—Hydroxy—3—Methylglutaryl Coenzyme A Reductase a Rate—Limiting Step for Isoprenoid Biosynthesis in Plants,” [21] Appl. No.: 631,341 Plant Physiol. 109:1337—1343 (1995). [22] Filed: Apr. 12, 1996 Chappell, “The Biochemistry and Molecular Biology of Isoprenoid Metabolism,” Plant Physiol. 107:1—6 (1995). [51] Int. Cl.6 ............................. .. C07K 5/00; c071< 7/00; Chen et al., “Cloning, Expression, and Characterization of c071< 17/00 (+)—6—Cadinene Synthase: A Catalyst for Cotton Phytoal [52] US. Cl. ........................ .. 530/350; 530/300; 530/324; exin Biosynthesis,” Arch. Biochem. Biophys. 324:255—266 530/370; 530/371; 530/372; 530/373; 530/375; (1995). 530/376; 530/377; 530/378; 530/379; 530/374; Chiu et al., “Engineered GFP as a vital reporter in plants,” 536/231; 536/234; 536/236; 536/2374; Current Biology 6:325—330 (1996), 1996. 435/252.1; 435/252.33; 435/320.1; 435/183; 435/69.1; 435/69.7; 435/71.1; 935/22; 935/31; Colby et al., “4S—Limonene Synthase from the Oil Glands of 935/52 Spearmint (Mentha spicata),” J. Biol. Chem. 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Cytotoxic Activities of TWenty Unsaturated Sesquiterpene Elakovich, “Sesquiterpenes as Phytoalexins and Allelo Dialdehydes from Plants and Mushrooms,” Planta Med. pathic Agents,” Ecology and Metabolism of Plant Lipids 57:344 (1991). Ave et al., “Aphid repellent sesquiterpenes in glandular 325:93—108 (1986). trichomes of Solanum berthaultii and S. tuberosum,” Ento Enzell et al., “Mass spectra of tobacco isoprenoids,” Mass mologia Experimentalis et Applicata 44:131—138 (1987). Spectrometry Rev. 3:395 (1984). Back et al., “Expression of a Plant Sesquiterpene Cyclase (List continued on next page.) Gene in Escherichia coli, ” Arch. Biochem. Biophys. 315:527—532 (1994). Primary Examiner—Sheela Huff Back et al. “Cloning and Bacterial Expression of a Sesquit Attorney, Agent, or Firm—Clark & Elbing LLP erpene Cyclase from Hyoscyamus muticus and Its Molecular Comparison to Related Terpene Cyclases,” J. Biol. Chem. [57] ABSTRACT 270:7375—7381 (1995). Barnby et al. “Effects of Azadirachtin on Levels of Ecdys Disclosed is a chimeric isoprenoid synthase polypeptide teroids and Prothoracicotropic Hormone—Like Activity in including a ?rst domain from a ?rst isoprenoid synthase Heliothis Virescens (Fabr.) Larvae,” J. Insect. Physiol. joined to a second domain from a second, heterologous 36:125—131 (1990). isoprenoid synthase, Whereby the chimeric isoprenoid syn Borman, “Scientists Mobilize to Increase Supply of Anti thase is capable of catalyzing the production of isoprenoid cancer Drug Taxol,” Chemical & Engineering News 11—18 reaction products that are not produced in the absence of the (1991). second domain of the second, heterologous isoprenoid syn BoWers et al., “Sesquiterpene Progenitor, Germacrene A: An thase. 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RuZicka, “The Isoprene Rule and the Biogenesis of Terpenic Joly et al., “Effect of Site—directed Mutagenesis of Con Compounds,” Experientia 10:357—396 (1953). served Asparate and Arginine Residues upon Farnesyl Savage et al., “Monoterpene Synthases of Pinus contorta Diphosphate Synthase Activity,” J. of Biol. Chem. and Related Conifers,” J. Biol. Chem. 269:4012—4020 268:26983—26989 (1993). Kalsi et al., “Stereostructures of TWo Biologically Active (1994). Sesquiterpene Lactones from Inula Racemosa, ” Phytochem Smith et al., “The Syringolides: Bacterial C—Glycosyl Lip istry 28:2093—2096 (1989). ids That Trigger Plant Disease Resistance,” Tetrahedron
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