DNA encoding limonene synthase from Mentha spicata

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for...

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435 691, 4352523, 4353201, 536 232, C12N 900, C12N 1563, C12P 2102, C07H 2104

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058719881

ABSTRACT:
cDNA encoding (-)-4S-limonene synthase from spearmint has been isolated and sequenced, and the corresponding amino acid sequence has been determined. Accordingly, isolated DNA sequences are provided which code for the expression of limonene synthase, such as the sequence designated SEQ ID No:11 which encodes limonene synthase from spearmint (Mentha spicata). In other aspects, replicable recombinant cloning vehicles are provided which code for limonene synthase or for a base sequence sufficiently complementary to at least a portion of the limonene synthase DNA or RNA to enable hybridization therewith (e.g., antisense limonene synthase RNA or fragments of complementary limonene synthase DNA which are useful as polymerase chain reaction primers or as probes for limonene synthase or related genes). In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding limonene synthase. Thus, systems and methods are provided for the recombinant expression of limonene synthase that may be used to facilitate the production, isolation and purification of significant quantities of recombinant limonene synthase (or of the primary enzyme product, limonene) for subsequent use, to obtain expression or enhanced expression of limonene synthase in plants to attain enhanced limonene production as a predator defense mechanism, or may be otherwise employed for the regulation or expression of limonene synthase or the production of limonene.

REFERENCES:
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John, M.E. "An efficient method for isolation of RNA and DNA from plants containing polyphenolics" Nucleic Acids Research (May, 1992), vol. 20, No. 9, p. 2381.
Alberts, B. et al. "Molecular Biology of the Cell" published 1989 by Garland Publishing Inc. (N.Y.), pp. 262-263.
Alberts, B. et al. (1989) Molecular Biology of the Cell Second Ed. Garland Publishing Inc., New York. pp. 185-187 and 265-266.
Colby, S.M., Alonso, W.R., Croteau, R. (1992) "Isolation and characterization of cDNA encoding limonene cyclase in spearmint" J. Cell Biochem. Suppl. 0 vol. 16, Part F p. 230.

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