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公开(公告)号：US08742203B2
公开(公告)日：2014-06-03
申请号：US12762941
申请日：2010-04-19
CPC分类号： C12N15/8247 , C12N15/8237 , C12N15/8243
摘要： Methods and constructs for the introduction of multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway are provided. In one embodiment, the constructs contain two or more enzyme-encoding genes, each under the control of an inducible promoter and each with a polyadenylation signal. The constructs are used to produce transgenic plants, in which the expression of the enzymes are increased when a chemical inducing agent is applied, and a biosynthetic product of the series of enzymes encoded by the transgenes is produced. Constructs may be used which contain two or more enzyme-encoding genes under the control of one or more promoters activated by activator molecules or complexes expressed from a transgene or transgenes, which are themselves under the control of one or more inducible promoters and switched on following the external application of a chemical. The transgene or transgenes expressing the activator molecules or complexes may be included in the same construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. Alternatively, the transgene or transgenes expressing the activator molecules or complexes may be on a different construct from the construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. The activator molecule can be expressed using a constitutive promoter in an inactive form which is converted to the active form following application of the chemical inducing agent.
摘要翻译： 提供了在多酶生物合成途径中引入编码酶的多个基因的方法和构建体。 在一个实施方案中，构建体含有两个或更多个酶编码基因，每个基因在诱导型启动子的控制下并且各自具有多聚腺苷酸化信号。 构建体用于产生转基因植物，其中当施用化学诱导剂时酶的表达增加，并且产生由转基因编码的一系列酶的生物合成产物。 可以使用含有两个或更多个酶编码基因的构建体，所述两个或多个酶编码基因在一个或多个由活化剂分子或由转基因或转基因表达的复合物所激活的启动子的控制下，所述启动子本身处于一个或多个诱导型启动子的控制下， 化学品的外部应用。 表达活化剂分子或复合物的转基因或转基因可以包含在含有多酶生物合成途径中的酶的多个基因的相同构建体中。 或者，表达活化剂分子或复合物的转基因或转基因可以与含有多酶生物合成途径中的酶的多个基因的构建体在不同的构建体上。 可以使用非活性形式的组成型启动子来表达活化剂分子，其在施用化学诱导剂后转化为活性形式。
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公开(公告)号：US20130042371A1
公开(公告)日：2013-02-14
申请号：US13590933
申请日：2012-08-21
CPC分类号： C12N15/8247 , C12N15/8237 , C12N15/8243
摘要： Methods and constructs for the introduction of multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway are provided. In one embodiment, the constructs contain two or more enzyme-encoding genes, each under the control of an inducible promoter and each with a polyadenylation signal. The constructs are used to produce transgenic plants, in which the expression of the enzymes are increased when a chemical inducing agent is applied, and a biosynthetic product of the series of enzymes encoded by the transgenes is produced. Constructs may be used which contain two or more enzyme-encoding genes under the control of one or more promoters activated by activator molecules or complexes expressed from a transgene or transgenes, which are themselves under the control of one or more inducible promoters and switched on following the external application of a chemical. The transgene or transgenes expressing the activator molecules or complexes may be included in the same construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. Alternatively, the transgene or transgenes expressing the activator molecules or complexes may be on a different construct from the construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. The activator molecule can be expressed using a constitutive promoter in an inactive form which is converted to the active form following application of the chemical inducing agent.
摘要翻译： 提供了在多酶生物合成途径中引入编码酶的多个基因的方法和构建体。 在一个实施方案中，构建体含有两个或更多个酶编码基因，每个基因在诱导型启动子的控制下并且各自具有多聚腺苷酸化信号。 构建体用于产生转基因植物，其中当施用化学诱导剂时酶的表达增加，并且产生由转基因编码的一系列酶的生物合成产物。 可以使用含有两个或更多个酶编码基因的构建体，所述两个或多个酶编码基因在一个或多个由活化剂分子或由转基因或转基因表达的复合物所激活的启动子的控制下，所述启动子本身处于一个或多个诱导型启动子的控制下， 化学品的外部应用。 表达活化剂分子或复合物的转基因或转基因可以包含在含有多酶生物合成途径中的酶的多个基因的相同构建体中。 或者，表达活化剂分子或复合物的转基因或转基因可以与含有多酶生物合成途径中的酶的多个基因的构建体在不同的构建体上。 可以使用非活性形式的组成型启动子来表达活化剂分子，其在施用化学诱导剂后转化为活性形式。
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公开(公告)号：US20100196974A1
公开(公告)日：2010-08-05
申请号：US12762941
申请日：2010-04-19
CPC分类号： C12N15/8247 , C12N15/8237 , C12N15/8243
摘要： Methods and constructs for the introduction of multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway are provided. In one embodiment, the constructs contain two or more enzyme-encoding genes, each under the control of an inducible promoter and each with a polyadenylation signal. The constructs are used to produce transgenic plants, in which the expression of the enzymes are increased when a chemical inducing agent is applied, and a biosynthetic product of the series of enzymes encoded by the transgenes is produced. Constructs may be used which contain two or more enzyme-encoding genes under the control of one or more promoters activated by activator molecules or complexes expressed from a transgene or transgenes, which are themselves under the control of one or more inducible promoters and switched on following the external application of a chemical. The transgene or transgenes expressing the activator molecules or complexes may be included in the same construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. Alternatively, the transgene or transgenes expressing the activator molecules or complexes may be on a different construct from the construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. The activator molecule can be expressed using a constitutive promoter in an inactive form which is converted to the active form following application of the chemical inducing agent.
摘要翻译： 提供了在多酶生物合成途径中引入编码酶的多个基因的方法和构建体。 在一个实施方案中，构建体含有两个或更多个酶编码基因，每个基因在诱导型启动子的控制下并且各自具有多聚腺苷酸化信号。 构建体用于产生转基因植物，其中当施用化学诱导剂时酶的表达增加，并且产生由转基因编码的一系列酶的生物合成产物。 可以使用含有两个或更多个酶编码基因的构建体，所述两个或多个酶编码基因在一个或多个由活化剂分子或由转基因或转基因表达的复合物所激活的启动子的控制下，所述启动子本身处于一个或多个诱导型启动子的控制下， 化学品的外部应用。 表达活化剂分子或复合物的转基因或转基因可以包含在含有多酶生物合成途径中的酶的多个基因的相同构建体中。 或者，表达活化剂分子或复合物的转基因或转基因可以与含有多酶生物合成途径中的酶的多个基因的构建体在不同的构建体上。 可以使用非活性形式的组成型启动子来表达活化剂分子，其在施用化学诱导剂后转化为活性形式。
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公开(公告)号：US07732680B2
公开(公告)日：2010-06-08
申请号：US11376831
申请日：2006-03-16
CPC分类号： C12N15/8247 , C12N15/8237 , C12N15/8243
摘要： Methods and constructs for the introduction of multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway are provided. In one embodiment, the constructs contain two or more enzyme-encoding genes, each under the control of an inducible promoter and each with a polyadenylation signal. The constructs are used to produce transgenic plants, in which the expression of the enzymes are increased when a chemical inducing agent is applied, and a biosynthetic product of the series of enzymes encoded by the transgenes is produced. Constructs may be used which contain two or more enzyme-encoding genes under the control of one or more promoters activated by activator molecules or complexes expressed from a transgene or transgenes, which are themselves under the control of one or more inducible promoters and switched on following the external application of a chemical. The transgene or transgenes expressing the activator molecules or complexes may be included in the same construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. Alternatively, the transgene or transgenes expressing the activator molecules or complexes may be on a different construct from the construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. The activator molecule can be expressed using a constitutive promoter in an inactive form which is converted to the active form following application of the chemical inducing agent.
摘要翻译： 提供了在多酶生物合成途径中引入编码酶的多个基因的方法和构建体。 在一个实施方案中，构建体含有两个或更多个酶编码基因，每个基因在诱导型启动子的控制下并且各自具有多聚腺苷酸化信号。 构建体用于产生转基因植物，其中当施用化学诱导剂时酶的表达增加，并且产生由转基因编码的一系列酶的生物合成产物。 可以使用含有两个或更多个酶编码基因的构建体，所述两个或多个酶编码基因在一个或多个由活化剂分子或由转基因或转基因表达的复合物所激活的启动子的控制下，所述启动子本身处于一个或多个诱导型启动子的控制下， 化学品的外部应用。 表达活化剂分子或复合物的转基因或转基因可以包含在含有多酶生物合成途径中的酶的多个基因的相同构建体中。 或者，表达活化剂分子或复合物的转基因或转基因可以与含有多酶生物合成途径中的酶的多个基因的构建体在不同的构建体上。 可以使用非活性形式的组成型启动子来表达活化剂分子，其在施用化学诱导剂后转化为活性形式。
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公开(公告)号：US20090271889A1
公开(公告)日：2009-10-29
申请号：US12431428
申请日：2009-04-28
申请人： Mariya N. Somleva , Kristi D. Snell , Julie Beaulieu , Oliver P. Peoples , Bradley Garrison , Nii Patterson
发明人： Mariya N. Somleva , Kristi D. Snell , Julie Beaulieu , Oliver P. Peoples , Bradley Garrison , Nii Patterson
CPC分类号： C12N9/1025 , C12N9/0004 , C12N9/1029 , C12N15/8243
摘要： Transgenic plants, plant material, and plant cells for synthesis of polyhydroxyalkanoates, preferably poly(3-hydroxybutyrate) (also referred to a as PHB) are provided. Preferred plants that can be genetically engineered to produce PHB include plants that do not normally produce storage products such as oils and carbohydrates, and plants that have a C4 NAD-malic enzyme photosynthetic pathway. Such plants also advantageously produce lignocellulosic biomass that can be converted into biofuels. An exemplary plant that can be genetically engineered to produce PHB and produce lignocellulosic biomass is switchgrass, Panicum virgatum L. A preferred cultivar of switchgrass is Alamo. Other suitable cultivars of switchgrass include but are not limited to Blackwell, Kanlow, Nebraska 28, Pathfinder, Cave-in-Rock, Shelter and Trailblazer.
摘要翻译： 提供用于合成聚羟基链烷酸酯，优选聚（3-羟基丁酸酯）（也称为PHB）的转基因植物，植物材料和植物细胞。 可以进行遗传工程以生产PHB的优选植物包括通常不产生储存产物如油和碳水化合物的植物，以及具有C4 NAD-苹果酸光合途径的植物。 这样的植物还有利地生产可以转化为生物燃料的木质纤维素生物质。 可以遗传工程化生产PHB并生产木质纤维素生物质的示范性植物是柳枝稷（Panicum virgatum L.）。优选的柳枝稷品种是Alamo。 柳枝稷的其他合适品种包括但不限于Blackwell，Kanlow，Nebraska 28，Pathfinder，Cave-in-Rock，Shelter和Trailblazer。
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公开(公告)号：US06586658B1
公开(公告)日：2003-07-01
申请号：US09389395
申请日：1999-09-03
IPC分类号： A01H100
CPC分类号： C12N15/8289 , C12N15/52 , C12N15/8243 , C12N15/8247
摘要： Methods and systems to modify fatty acid biosynthesis and oxidation in plants to make new polymers are provided. Two enzymes are essential: a hydratase such as D-specific enoyl-CoA hydratase, for example, the hydratase obtained from Aeromonas caviae, and a &bgr;-oxidation enzyme system. Some plants have a &bgr;-oxidation enzyme system which is sufficient to modify polymer synthesis when the plants are engineered to express the hydratase. Examples demonstrate production of polymer by expression of these enzymes in transgenic plants. Examples also demonstrate that modifications in fatty acid biosynthesis can be used to alter plant phenotypes, decreasing or eliminating seed production and increasing green plant biomass, as well as producing polyhydroxyalkanoates.
摘要翻译： 提供了修饰植物脂肪酸生物合成和氧化以制备新聚合物的方法和系统。 两种酶是必需的：水合酶如D-特异性烯酰辅酶A水合酶，例如从气单胞菌（Aeromonas caviae）获得的水合酶和β-氧化酶系统。 一些植物具有β-氧化酶系统，当植物被工程化以表达水合酶时，其足以改变聚合物合成。 实施例证明通过在转基因植物中表达这些酶来生产聚合物。 实例还表明，脂肪酸生物合成中的修饰可用于改变植物表型，减少或消除种子生产和增加绿色植物生物量，以及生产聚羟基链烷酸酯。
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7.发明申请STABLE, FERTILE, HIGH POLYHYDROXYALKANOATE PRODUCING PLANTS AND METHODS OF PRODUCING THEM 有权稳定，有效的高聚羟基烷基酯生产植物及其生产方法公开(公告)号：US20100229258A1
公开(公告)日：2010-09-09
申请号：US12718498
申请日：2010-03-05
CPC分类号： C12N15/8214 , C12N15/8257
摘要： Transgenic plants that produce high levels of polyhydroxybutyrate and methods of producing them are provided. In a preferred embodiment the transgenic plants are produced using plastid transformation technologies and utilize genes which are codon optimized. Stably transformed plants able to produce greater than 10% dwt PHS in tissues are also provided.
摘要翻译： 提供了产生高水平聚羟基丁酸的转基因植物及其生产方法。 在优选的实施方案中，使用质体转化技术产生转基因植物并利用密码子优化的基因。 还提供了能够在组织中产生大于10％dwt PHS的稳定转化的植物。
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公开(公告)号：US09181559B2
公开(公告)日：2015-11-10
申请号：US13395702
申请日：2010-09-15
申请人： Nii Patterson , Jihong Tang , Jixiang Han , Venkata Tavva , Andrew Hertig , Zhigang Zhang , Thomas Martin Ramseier , Karen Bohmert-Tatarev , Oliver P. Peoples , Kristi D. Snell
发明人： Nii Patterson , Jihong Tang , Jixiang Han , Venkata Tavva , Andrew Hertig , Zhigang Zhang , Thomas Martin Ramseier , Karen Bohmert-Tatarev , Oliver P. Peoples , Kristi D. Snell
CPC分类号： C12N15/8214 , C12N15/8257
摘要： Transgenic plants, plant material, plant cells, and genetic constructs for synthesis of biopolymers, for example polyhydroxyalkanoates (“PHA”) are provided. In one embodiment, the transgenic plants synthesize polyhydroxybutyrate (“PHB”). In one embodiment the transgenic plant encodes siRNA for one or more of the genes encoding enzymes for producing PHA. In a more preferred embodiment, the siRNA expression is under the control of an inducible regulatory element. In another embodiment, the transgenic plant contains transgenes that encode expression enzymes that will degrade the polymer. In a preferred embodiment, the expression of these enzymes is under the control of a germination specific, inducible, or minimal promoter. In another embodiment, the transgenic plant contains transgenes encoding enzymes that increase carbon flow for polymer synthesis. In a preferred embodiment, these transgenes encode enzymes that increase carbon flow in the Calvin Cycle.
摘要翻译： 提供了用于合成生物聚合物的转基因植物，植物材料，植物细胞和遗传构建体，例如聚羟基链烷酸酯（“PHA”）。 在一个实施方案中，转基因植物合成聚羟基丁酸酯（“PHB”）。 在一个实施方案中，转基因植物编码编码用于产生PHA的酶的一种或多种基因的siRNA。 在更优选的实施方案中，siRNA表达受诱导型调节元件的控制。 在另一个实施方案中，转基因植物含有编码将降解聚合物的表达酶的转基因。 在优选的实施方案中，这些酶的表达受发芽特异性，诱导型或最小启动子的控制。 在另一个实施方案中，转基因植物含有编码增加聚合物合成碳流的酶的转基因。 在优选的实施方案中，这些转基因编码增加卡尔文循环中碳流的酶。
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9.发明授权Production of medium chain length polyhydroxyalkanoates from fatty acid biosynthetic pathways 有权从脂肪酸生物合成途径生产中链长度聚羟基链烷酸酯公开(公告)号：US07786355B2
公开(公告)日：2010-08-31
申请号：US09991152
申请日：2001-11-16
CPC分类号： C12N9/93 , C12N9/00 , C12N9/10 , C12N9/1029 , C12N9/16 , C12N15/8243
摘要： Methods for producing polyhydroxyalkanoates (PHAs) from fatty acid biosynthetic pathways using a 3-hydroxy acyl ACP thioesterase, a PHA synthase, and an acyl CoA synthetase, have been developed. Methodology for enabling PHA production from fatty acid biosynthetic pathways in non-native bacterial PHA producers and plants using an enzyme having the catalytic activity of 3-hydroxy acyl ACP thioesterase, an acyl CoA synthetase with substrate specificity for medium chain length 3-hydroxy fatty acids, and a medium chain length PHA synthase, has been developed. Acyl CoA synthetase activity can be supplied either by the endogenous acyl CoA synthetase of the host organism, when sufficiently expressed, or the host organism's activity can be supplemented by the expression of a recombinant acyl CoA synthetase gene. New strategies are described for plant based PHA production in the chloroplasts, cytosol, and peroxisomes of biomass crops as well as the plastids, cytosol, and peroxisomes of oil seed crops.
摘要翻译： 已经开发了使用3-羟基酰基ACP硫酯酶，PHA合成酶和酰基辅酶A合成酶从脂肪酸生物合成途径制备聚羟基链烷酸酯（PHA）的方法。 使用具有3-羟基酰基ACP硫酯酶的催化活性的酶，非天然细菌PHA生产者和植物中的脂肪酸生物合成途径的PHA生产的方法，具有对于中链长度3-羟基脂肪酸具有底物特异性的酰基辅酶A合成酶 ，和中链长度PHA合酶。 酰基辅酶A合成酶活性可以通过宿主生物的内源酰基辅酶A合成酶提供，当充分表达时，或者宿主生物的活性可以通过重组酰基辅酶A合成酶基因的表达来补充。 描述了用于生物量作物的叶绿体，细胞溶质和过氧化物酶体以及油料种子作物的质体，细胞溶质和过氧化物酶体的基于植物的PHA生产的新策略。
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公开(公告)号：US20120180162A1
公开(公告)日：2012-07-12
申请号：US13395702
申请日：2010-09-15
申请人： Nii Patterson , Jihong Tang , Jixiang Han , Venkata Tavva , Andrew Hertig , Zhigang Zhang , Thomas Martin Ramseier , Karen Bohmert-tatarev , Oliver P. peoples , Kristi D. Snell
发明人： Nii Patterson , Jihong Tang , Jixiang Han , Venkata Tavva , Andrew Hertig , Zhigang Zhang , Thomas Martin Ramseier , Karen Bohmert-tatarev , Oliver P. peoples , Kristi D. Snell
CPC分类号： C12N15/8214 , C12N15/8257
摘要： Transgenic plants, plant material, plant cells, and genetic constructs for synthesis of biopolymers, for example polyhydroxyalkanoates (“PHA”) are provided. In one embodiment, the transgenic plants synthesize polyhydroxybutyrate (“PHB”). In one embodiment the transgenic plant encodes siRNA for one or more of the genes encoding enzymes for producing PHA. In a more preferred embodiment, the siRNA expression is under the control of an inducible regulatory element. In another embodiment, the transgenic plant contains transgenes that encode expression enzymes that will degrade the polymer. In a preferred embodiment, the expression of these enzymes is under the control of a germination specific, inducible, or minimal promoter. In another embodiment, the transgenic plant contains transgenes encoding enzymes that increase carbon flow for polymer synthesis. In a preferred embodiment, these transgenes encode enzymes that increase carbon flow in the Calvin Cycle.
摘要翻译： 提供了用于合成生物聚合物的转基因植物，植物材料，植物细胞和遗传构建体，例如聚羟基链烷酸酯（“PHA”）。 在一个实施方案中，转基因植物合成聚羟基丁酸酯（“PHB”）。 在一个实施方案中，转基因植物编码编码用于产生PHA的酶的一种或多种基因的siRNA。 在更优选的实施方案中，siRNA表达受诱导型调节元件的控制。 在另一个实施方案中，转基因植物含有编码将降解聚合物的表达酶的转基因。 在优选的实施方案中，这些酶的表达受发芽特异性，诱导型或最小启动子的控制。 在另一个实施方案中，转基因植物含有编码增加聚合物合成碳流的酶的转基因。 在优选的实施方案中，这些转基因编码增加卡尔文循环中碳流的酶。
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