SESQUITERPENE SYNTHASES FROM GRAND FIR ABIES GRANDIS, AND METHODS OF USE

摘要

cDNAs encoding E-.alpha.-bisabolene synthase, .delta.-selinene synthase and.gamma.-humulene synthase from Grand Fir (Abies grandis) have been isolatedand sequenced, and the corresponding amino acid sequences have beendetermined. Accordingly, isolated DNA sequences (SEQ ID No:12; SEQ ID No:19and SEQ ID No:23) are provided which code for the expression of E-.alpha.-bisabolene synthase (SEQ ID No:13), .delta.-selinene synthase (SEQ ID No:20)and .gamma.-humulene synthase (SEQ ID No:24), respectively, from Grand Fir(Abies grandis). In other aspects, replicable recombinant cloning vehicles areprovided which code for E-.alpha.-bisabolene synthase, .delta.-selinenesynthase and .gamma.-humulene synthase, or for a base sequence sufficientlycomplementary to at least a portion of E-.alpha.-bisabolene synthase, .delta.-selinene synthase or .gamma.-humulene synthase DNA or RNA to enablehybridization therewith. In yet other aspects, modified host cells areprovided that have been transformed, transfected, infected and/or injectedwith a recombinant cloning vehicle and/or DNA sequence encoding E-.alpha.-bisabolene synthase, .delta.-selinene synthase or .gamma.-humulene synthase.Thus, systems and methods are provided for the recombinant expression of theaforementioned recombinant sesquiterpene synthases that may be used tofacilitate their production, isolation and purification in significantamounts. Recombinant E-.alpha.-bisabolene synthase, .delta.-selinene synthaseand .gamma.-humulene synthase may be used to obtain expression or enhancedexpression of E-.alpha.-bisabolene synthase, .delta.-selinene synthase and.gamma.-humulene synthase in plants in order to enhance the production ofsesquiterpenoids, or may be otherwise employed for the regulation orexpression of E-.alpha.-bisabolene synthase, .delta.-selinene synthase and.gamma.-humulene synthase, or the production of their products.