公开(公告)号：US20020038001A1

公开(公告)日：2002-03-28

申请号：US09865171

申请日：2001-05-24

Applicant: Washington State University Research Foundation

Inventor： Rodney Bruce Croteau ,  Jorg Bohlmann ,  John E. Crock ,  Christopher L. Steele

IPC: C12P021/00

CPC classification number: C12N15/8243 ,  C12N9/88 ,  C12N15/8279

Abstract: cDNAs encoding E-null-bisabolelene synthase, null-selinene synthase and null-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-null-bisabolene synthase (SEQ ID No:13), null-selinene synthase (SEQ ID No:20) and null-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-null-bisabolene synthase, null-selinene synthase and null-humulene synthase, or for a base sequence sufficiently complementary to at least a portion of E-null-bisabolene synthase, null-selinene synthase or null-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-null-bisabolene synthase, null-selinene synthase or null-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-null-bisabolene synthase, null-selinene synthase and null-humulene synthase may be used to obtain expression or enhanced expression of E-null-bisabolene synthase, null-selinene synthase and null-humulene synthase in plants in order to enhance the production of sesquiterpenoids, or may be otherwise employed for the regulation or expression of E-null-bisabolene synthase, null-selinene synthase and null-humulene synthase, or the production of their products.

公开(公告)号：US20030166204A1

公开(公告)日：2003-09-04

申请号：US09860282

申请日：2001-05-17

Applicant: Washington State University Research Foundation

Inventor： Rodney Bruce Croteau ,  John E. Crock

IPC: C12N009/10 ,  C07H021/04 ,  C12P021/02 ,  C12N005/04

CPC classification number: C12P5/007 ,  C12N9/88

Abstract: A cDNA encoding (E)-null-farnesene synthase from peppermint (Mentha piperita) has been isolated and sequenced, and the corresponding amino acid sequence has been determined. Accordingly, an isolated DNA sequence (SEQ ID NO:1) is provided which codes for the expression of (E)-null-farnesene synthase (SEQ ID NO:2), from peppermint (Mentha piperisa). In other aspects, replicable recombinant cloning vehicles are provided which code for (E)-null-farnesene synthase, or for a base sequence sufficiently complementary to at least a portion of (E)-null-farnesene 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)-null-farnesene synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant (E)-null-famesene synthase that may be used to facilitate its production, isolation and purification in significant amounts. Recombinant (E)-null-farnesene synthase may be used to obtain expression or enhanced expression of (E)-null-famesene synthase in plants in order to enhance the production of (E)-null-farnesene, or may be otherwise employed for the regulation or expression of (E)-null-farnesene synthase, or the production of its product.

Abstract translation: 已经分离了来自薄荷（薄荷胡椒）的编码（E） - 法呢烯合成酶的cDNA并测序，并确定了相应的氨基酸序列。 因此，提供了从薄荷（Mentha piperisa）编码（E） - 法呢烯合成酶（SEQ ID NO：2）的表达的分离的DNA序列（SEQ ID NO：1）。 在其它方面，提供了编码（E） - 法呢烯合成酶的可重复的重组克隆载体，或与（E） - 法呢烯合成酶DNA或RNA的至少一部分充分互补的碱基序列，以使其能够与其进行杂交 。 在另一方面，提供已经用编码（E） - 法呢烯合成酶的重组克隆载体和/或DNA序列转化，转染，感染和/或注射的修饰的宿主细胞。 因此，提供了用于重组表达上述重组（E）-β-甲酰基合成酶的系统和方法，其可用于促进其显着量的产生，分离和纯化。 可以使用重组（E） - 法呢烯合成酶来获得植物中（E）-β-fameene合酶的表达或增强的表达，以增强（E） - 法呢烯的生产，或者可以另外用于 （E） - 法呢烯合成酶的调节或表达，或其产物的生产。