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SESQUITERPENE SYNTHASES FROM GRAND FIR $i(ABIES GRANDIS), AND METHODS OF USE

机译:大冷杉$ i(ABIES GRANDIS)的倍半萜烯合酶及其使用方法

摘要

cDNAs encoding E-α-bisabolene synthase, δ-selinene synthase and η-humulene synthase from Grand Fir (Abies grandis) have been isolated and sequenced, and the corresponding amino acid sequences have been determined. Accordingly, isolated DNA sequences (SEQ ID No:12; SEQ ID No:19 and SEQ ID No:23) are provided which code for the expression of E-α-bisabolene synthase (SEQ ID No:13), δ-selinene synthase (SEQ ID No:20) and η-humulene synthase (SEQ ID No:24), respectively, from Grand Fir (Abies grandis). In other aspects, replicable recombinant cloning vehicles are provided which code for E-α-bisabolene synthase, δ-selinene synthase and η-humulene synthase, or for a base sequence sufficiently complementary to at least a portion of E-α-bisabolene synthase, δ-selinene synthase or η-humulene synthase DNA or RNA to enable hybridization therewith. 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 E-α-bisabolene synthase, δ-selinene synthase or η-humulene synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant sesquiterpene synthases that may be used to facilitate their production, isolation and purification in significant amounts. Recombinant E-α-bisabolene synthase, δ-selinene synthase and η-humulene synthase may be used to obtain expression or enhanced expression of E-α-bisabolene synthase, δ-selinene synthase and η-humulene synthase in plants in order to enhance the production of sesquiterpenoids, or may be otherwise employed for the regulation or expression of E-α-bisabolene synthase, δ-selinene synthase and η-humulene synthase, or the production of their products.
机译:已分离并测序了来自Grand Fir(Abies grandis)的编码E-α-双-abobolene合酶,δ-selinene合酶和η-humulene合酶的cDNA,并确定了相应的氨基酸序列。因此,提供了分离的DNA序列(SEQ ID No:12; SEQ ID No:19和SEQ ID No:23),其编码E-α-双水杨烯合酶(SEQ ID No:13),δ-selinene合酶的表达。分别来自Grand Fir(Abies grandis)(SEQ ID No:20)和η-humulene合酶(SEQ ID No:24)。在其他方面,提供了可复制的重组克隆载体,其编码E-α-双水杨烯合酶,δ-selinene合酶和η-humulene合酶,或与至少一部分E-α-二水杨烯合酶充分互补的碱基序列, δ-selinene合酶或η-humulene合酶DNA或RNA使其能够杂交。在其他方面,提供了经修饰的宿主细胞,其已用重组克隆载体和/或编码E-α-双水杨烯合酶,δ-selinene合酶或η-humulene合酶的DNA序列进行转化,转染,感染和/或注射。因此,提供了用于上述重组人倍半萜合酶的重组表达的系统和方法,其可用于促进其大量生产,分离和纯化。重组E-α-bisabolene合酶,δ-selinene合酶和η-humulene合酶可用于在植物中表达或增强E-α-bisabolene合酶,δ-selinene合酶和η-humulene合酶的表达倍半萜类化合物的生产,或另外可用于调节或表达E-α-双水杨烯合酶,δ-selinene合酶和η-humulene合酶或其产物的生产。

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