公开(公告)号：US07067726B2

公开(公告)日：2006-06-27

申请号：US10455915

申请日：2003-06-06

申请人： Elizabeth Hood ,  John Howard ,  Richard Clough ,  Kameshwari Pappu

发明人： Elizabeth Hood ,  John Howard ,  Richard Clough ,  Kameshwari Pappu

IPC分类号： A01H5/00 ,  C12N15/82 ,  C12N15/87 ,  C12N5/02 ,  C07H21/04

CPC分类号： C12N9/0065 ,  C12N15/8257 ,  C12Y111/01013

摘要： Expression of a recombinant manganese-dependent peroxidase in plants is described. Methods for the recovery of transgenic corn seeds that express high levels of functional manganese-dependent peroxidase are also provided. Manganese-dependent peroxidase vectors are engineered to contain sequences directing seed-preferred expression as well as sequences for cell wall-targeted localization.

摘要翻译： 描述了重组锰依赖性过氧化物酶在植物中的表达。 还提供了回收表达高水平功能性锰依赖性过氧化物酶的转基因玉米种子的方法。 锰依赖性过氧化物酶载体被工程化以含有指导种子优选表达的序列以及用于细胞壁靶向定位的序列。

公开(公告)号：US07541515B2

公开(公告)日：2009-06-02

申请号：US10252732

申请日：2002-09-23

申请人： Elizabeth Hood ,  John Howard ,  Donna Delaney

发明人： Elizabeth Hood ,  John Howard ,  Donna Delaney

IPC分类号： C12N15/82

CPC分类号： C12N15/8216 ,  C12N15/8257

摘要： Increased expression of heterologous proteins in a plant is achieved by introducing the nucleotide sequence encoding the heterologous protein into a plant that has high oil seed content compared to low oil seed and/or is introduced into a plant having reduced alcohol soluble proteins in the endosperm. The nucleotide sequence may be introduced by direct transformation into the plant, or by direct transformation into another plant, and crossing with the high oil plant or plant have reduced levels of alcohol soluble protein in the endosperm. Further, the nucleotide sequences may be introduced into one or both of the high oil plant and the plant having reduced levels of alcohol soluble proteins, and the plants may be crossed to result in a progeny having even further increased expression levels of the heterologous protein.

摘要翻译： 通过将编码异源蛋白质的核苷酸序列引入与低油种子相比具有高油种子含量的植物和/或被引入具有在胚乳中具有降低的可溶性蛋白质的植物中来实现植物中异源蛋白质的表达增加。 核苷酸序列可以通过直接转化到植物中，或通过直接转化到另一种植物中，并与高油植物或植物杂交在胚乳中具有降低的可溶性蛋白质水平。 此外，可以将核苷酸序列引入高油植物和具有降低的可溶性蛋白质水平的植物中的一个或两个中，并且可以使植物杂交以产生具有甚至进一步增加异源蛋白质表达水平的后代。

公开(公告)号：US20060026715A1

公开(公告)日：2006-02-02

申请号：US11219180

申请日：2005-09-02

申请人： Elizabeth Hood ,  John Howard

发明人： Elizabeth Hood ,  John Howard

IPC分类号： A01H1/00 ,  A01H5/00 ,  C12N9/24 ,  C12N5/04 ,  C12N15/82

CPC分类号： C12N15/8242 ,  C12N15/8246 ,  C12N15/8257

摘要： Expression of recombinant polysaccharide degrading enzymes in plants is described. In one embodiment, expression of the enzyme is preferentially directed to the seed of the plant. Expression may also be preferentially targeted to specific locations within the plant cell. Expression of cellulases in corn is shown. The result is the capacity to produce polysaccharide degrading enzymes in plants at commercially acceptable levels in a reliable manner. Methods of using same in production of ethanol is also described, including use of the plant-produced enzymes in the ethanol production process.

公开(公告)号：US20060015971A1

公开(公告)日：2006-01-19

申请号：US11226788

申请日：2005-09-14

申请人： John Howard ,  Elizabeth Hood

发明人： John Howard ,  Elizabeth Hood

IPC分类号： A01H1/00 ,  C12N15/82

CPC分类号： C12N9/6427 ,  C12N15/8257

摘要： Production of proteases in plants is set forth, whereby heterologous DNA encoding the protease is introduced into the plant and expression of the protein achieved. By such methods, expression is achieved in plants wherein the plant cell is not damaged, the protein can be recovered without contamination by other proteases, and can be expressed at levels such that commercial production of the enzyme is obtained. Expression levels can be at 0.1% of total soluble protein of the plant, or higher.

摘要翻译： 阐述植物中蛋白酶的产生，由此将编码蛋白酶的异源DNA引入植物中并实现蛋白质的表达。 通过这样的方法，在植物细胞未受损害的植物中实现表达，可以在不受其它蛋白酶污染的情况下回收蛋白质，并且可以以获得酶的商业生产的水平来表达。 表达水平可以在植物总可溶性蛋白质的0.1％以上。

公开(公告)号：US08558058B2

公开(公告)日：2013-10-15

申请号：US11219180

申请日：2005-09-02

申请人： Elizabeth Hood ,  John Howard

发明人： Elizabeth Hood ,  John Howard

IPC分类号： C12N15/82 ,  A01H5/00

CPC分类号： C12N15/8242 ,  C12N15/8246 ,  C12N15/8257

摘要： Expression of recombinant polysaccharide degrading enzymes in plants is described. In one embodiment, expression of the enzyme is preferentially directed to the seed of the plant. Expression may also be preferentially targeted to specific locations within the plant cell. Expression of cellulases in corn is shown. The result is the capacity to produce polysaccharide degrading enzymes in plants at commercially acceptable levels in a reliable manner. Methods of using same in production of ethanol is also described, including use of the plant-produced enzymes in the ethanol production process.

摘要翻译： 描述了重组多糖降解酶在植物中的表达。 在一个实施方案中，酶的表达优先针对植物的种子。 表达也可以优先靶向植物细胞内的特定位置。 显示玉米纤维素酶的表达。 结果是以可靠的方式以商业上可接受的水平在植物中产生多糖降解酶的能力。 还描述了在乙醇生产中使用其的方法，包括在乙醇生产过程中使用植物生产的酶。

公开(公告)号：US07981442B2

公开(公告)日：2011-07-19

申请号：US11427034

申请日：2006-06-28

申请人： Elizabeth Hood ,  Joel A. Strom ,  Michael VanAuker

发明人： Elizabeth Hood ,  Joel A. Strom ,  Michael VanAuker

IPC分类号： A61K9/127 ,  A61K9/48

CPC分类号： A61K9/1272 ,  A61K9/0009 ,  A61K41/0028 ,  A61K49/0002

摘要： A method of targeted drug delivery and imaging using nonionic surfactant vesicles (niosomes) in combination with ultrasound. Niosomes have potential applications in targeted drug delivery and imaging because of their ability to encapsulate therapeutic agents and their enhanced uptake by physiological membranes. Ultrasound may be used to mediate delivery non-invasively by altering the niosome membrane structure. Niosomes composed of polyoxyethylene sorbitan monostearate (Tween 61), cholesterol, and dicetyl phosphate were synthesized via a thin film hydration technique and used for encapsulation studies. Carboxyfluorescein dye (CF) was used as a drug model to demonstrate delivery. The amount of dye in the niosomes, the concentration of the vesicles, and their mean particle size after each 5 minute incremental exposure to ultrasound was monitored. Dye concentration in niosome samples decreased while the population and size distribution of the niosome remained largely unchanged. Ultrasound is demonstrated to enhance the rate of dye diffusion across the niosome membrane non-destructively.

摘要翻译： 使用非离子表面活性剂囊泡（niosomes）与超声波组合的靶向药物递送和成像的方法。 Niosomes由于其能够包封治疗剂和增强生理膜吸收的能力，在靶向药物递送和成像中具有潜在应用。 超声波可用于通过改变niosome膜结构非侵入性地介导递送。 通过薄膜水合技术合成由聚氧乙烯脱水山梨醇单硬脂酸酯（吐温61），胆固醇和磷酸二鲸蜡酯组成的纳米粒子，用于封装研究。 使用羧基荧光素染料（CF）作为药物模型来证明递送。 监测niosomes中的染料量，囊泡浓度及每5分钟增加暴露于超声波后的平均粒径。 niosome样品中的染料浓度下降，而niosome的人口和大小分布基本保持不变。 证明超声可以非破坏性地提高染色体在染色体膜上的扩散速率。

公开(公告)号：US20060292211A1

公开(公告)日：2006-12-28

申请号：US11427034

申请日：2006-06-28

申请人： Elizabeth Hood ,  Joel Strom ,  Michael VanAuker

发明人： Elizabeth Hood ,  Joel Strom ,  Michael VanAuker

IPC分类号： A61K9/127

CPC分类号： A61K9/1272 ,  A61K9/0009 ,  A61K41/0028 ,  A61K49/0002

摘要： A method of targeted drug delivery and imaging using nonionic surfactant vesicles (niosomes) in combination with ultrasound. Niosomes have potential applications in targeted drug delivery and imaging because of their ability to encapsulate therapeutic agents and their enhanced uptake by physiological membranes. Ultrasound may be used to mediate delivery non-invasively by altering the niosome membrane structure. Niosomes composed of polyoxyethylene sorbitan monostearate (Tween 61), cholesterol, and dicetyl phosphate were synthesized via a thin film hydration technique and used for encapsulation studies. Carboxyfluorescein dye (CF) was used as a drug model to demonstrate delivery. The amount of dye in the niosomes, the concentration of the vesicles, and their mean particle size after each 5 minute incremental exposure to ultrasound was monitored. Dye concentration in niosome samples decreased while the population and size distribution of the niosome remained largely unchanged. Ultrasound is demonstrated to enhance the rate of dye diffusion across the niosome membrane non-destructively.

摘要翻译： 使用非离子表面活性剂囊泡（niosomes）与超声波组合的靶向药物递送和成像的方法。 Niosomes由于其能够包封治疗剂和增强生理膜吸收的能力，在靶向药物递送和成像中具有潜在应用。 超声波可用于通过改变niosome膜结构非侵入性地介导递送。 通过薄膜水合技术合成由聚氧乙烯脱水山梨醇单硬脂酸酯（吐温61），胆固醇和磷酸二鲸蜡酯组成的纳米粒子，用于封装研究。 使用羧基荧光素染料（CF）作为药物模型来证明递送。 监测niosomes中的染料量，囊泡浓度及每5分钟增加暴露于超声波后的平均粒径。 niosome样品中的染料浓度下降，而niosome的人口和大小分布基本保持不变。 证明超声可以非破坏性地提高染色体在染色体膜上的扩散速率。

公开(公告)号：US6087558A

公开(公告)日：2000-07-11

申请号：US120582

申请日：1998-07-22

申请人： John A. Howard ,  Elizabeth Hood

发明人： John A. Howard ,  Elizabeth Hood

IPC分类号： C12N9/76 ,  C12N15/55 ,  C12N15/82 ,  A01H5/10 ,  A01H5/00 ,  C12N5/04 ,  C12N15/12 ,  C12N15/57

CPC分类号： C12N9/6427 ,  C12N15/8257

摘要： Production of proteases in plants is set forth, whereby heterologous DNA encoding the protease is introduced into the plant and expression of the protein achieved. By such methods, expression is achieved in plants wherein the plant cell is not damaged, the protein can be recovered without contamination by other proteases, and can be expressed at levels such that commercial production of the enzyme is obtained. Expression levels can be at 0.1% of total soluble protein of the plant, or higher.

摘要翻译： 阐述植物中蛋白酶的产生，由此将编码蛋白酶的异源DNA引入植物中并实现蛋白质的表达。 通过这样的方法，在植物细胞未受损害的植物中实现表达，可以在不受其它蛋白酶污染的情况下回收蛋白质，并且可以以获得酶的商业生产的水平来表达。 表达水平可以在植物总可溶性蛋白质的0.1％以上。

公开(公告)号：US20070172520A1

公开(公告)日：2007-07-26

申请号：US11561639

申请日：2006-11-20

申请人： Michael VanAuker ,  Anna Plaas ,  Elizabeth Hood

发明人： Michael VanAuker ,  Anna Plaas ,  Elizabeth Hood

IPC分类号： A61K39/395 ,  A61K9/127

CPC分类号： A61K39/00 ,  A61K9/1272 ,  A61K39/44 ,  A61K47/6913

摘要： An immunoniosmes for targeted delivery of therapeutic agents to specific tissues in a host and methods of synthesis of those niosomes. An antibody molecule having specificity for a target antigen, such as a cell surface marker or other marker differentially expressed on a target cell, is covalently coupled to a functionalized membrane constituent. In a particular embodiment the functionalized membrane constituent is polyoxyethylene sorbitan monostearate functionalized with cyanuric chloride. The niosomes of this invention thus provide a composition that enhances internalization or retention of the bioactive agent of the niosome into the cytoplasm of the cells of the target tissue by providing a high degree of target specificity. Furthermore, the membrane vesicle enhances the life of the therapeutic agent by preventing its degradation in the extracellular environment, while exhibiting lower toxicity than can occur with some liposomes. The niosomes of the present invention are thus particularly useful as vehicles for the delivery of therapeutics to specific target cells.

摘要翻译： 将治疗剂靶向递送到宿主中的特定组织的免疫组织和合成这些niosomes的方法。 对靶抗原（例如细胞表面标志物或靶细胞上差异表达的其他标记物）具有特异性的抗体分子与官能化的膜成分共价偶联。 在一个具体实施方案中，官能化膜组分是用氰尿酰氯官能化的聚氧乙烯脱水山梨醇单硬脂酸酯。 因此，本发明的niosomes提供了一种组合物，其通过提供高度的靶特异性来增强niosome的生物活性剂的内化或保留到靶组织的细胞的细胞质中。 此外，膜囊泡通过防止其在细胞外环境中的降解来增强治疗剂的寿命，同时表现出比一些脂质体可能发生的毒性更低的毒性。 因此，本发明的niosomes特别可用作将治疗剂递送到特定靶细胞的载体。

公开(公告)号：US5767379A

公开(公告)日：1998-06-16

申请号：US554586

申请日：1995-11-06

申请人： Chris Baszczynski ,  Elizabeth Hood ,  Sheila Maddock ,  Terry EuClaire Meyer ,  James C. Register, III ,  Derrick Witcher ,  John A. Howard

发明人： Chris Baszczynski ,  Elizabeth Hood ,  Sheila Maddock ,  Terry EuClaire Meyer ,  James C. Register, III ,  Derrick Witcher ,  John A. Howard

IPC分类号： C07K14/46 ,  C07K14/465 ,  C12N15/12 ,  C12N15/82 ,  A01H5/00 ,  C12N15/29

CPC分类号： C07K14/465 ,  C07K14/46 ,  C12N15/8257

摘要： A method for commercial production of avidin entails heterologous expression of the protein in plants, in native conformation, at an expression level such that avidin represents at least 0.1% of total extracted protein. A genetic map of the integration locus allows for the identification of the avidin-expressing plant. Genetic loci on a plant chromosome are revealed that support high levels of avidin expression and that can be used as a site of integration for high level expression of other genes of interest.

摘要翻译： 用于商业生产抗生物素蛋白的方法需要以天然构象在植物中异源表达蛋白质，其表达水平使得抗生物素蛋白占总提取蛋白质的至少0.1％。 整合位点的遗传图可以鉴定表达抗生物素蛋白的植物。 显示植物染色体上的遗传基因座，其支持高水平的抗生物素蛋白表达，并且可以用作其它感兴趣的基因的高水平表达的整合位点。