公开(公告)号：US5824870A

公开(公告)日：1998-10-20

申请号：US554161

申请日：1995-11-06

申请人： Chris Baszczynski ,  Thomas Czapla ,  Elizabeth Hood ,  Terry EuClaire Meyer ,  David Peterson ,  A. Gururaj Rao ,  James C. Register, III ,  Derrick Witcher ,  John A. Howard

发明人： Chris Baszczynski ,  Thomas Czapla ,  Elizabeth Hood ,  Terry EuClaire Meyer ,  David Peterson ,  A. Gururaj Rao ,  James C. Register, III ,  Derrick Witcher ,  John A. Howard

IPC分类号： C07K14/81 ,  C12N15/15 ,  C12N15/29 ,  C12N15/82 ,  C12N15/12 ,  A01H5/00

CPC分类号： C07K14/8117 ,  C12N15/8257 ,  C12N15/8286

摘要： A method for commercial production of aprotinin entails heterologous expression of the protein in plants, preferably at a level such that aprotinin accounts for at least 0.1% of the total extracted protein. An aprotinin-expressing plant also has potential of increased insecticidal resistance by virtue of producing the protein. A genetic map of the integration locus allows identification of plants derived from the transgenic plant. This approach also reveals genetic loci on a plant chromosome that support high levels of gene expression and can be used as site of integration for expression of other genes of interest.

摘要翻译： 抑肽酶的商业生产方法需要在植物中异源表达蛋白质，优选地使得抑肽酶占总提取蛋白质的至少0.1％。 表达抑肽酶的植物还具有通过产生蛋白质而具有增加的杀虫抗性的潜力。 整合位点的遗传图可以鉴定来自转基因植物的植物。 这种方法还揭示了支持高水平基因表达的植物染色体上的遗传基因座，并且可以用作其他感兴趣的基因表达的整合位点。

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。

公开(公告)号：US5804694A

公开(公告)日：1998-09-08

申请号：US554169

申请日：1995-11-06

申请人： Wesley B. Bruce ,  Elizabeth Hood ,  David J. Peterson ,  James C. Register, III ,  Derrick Witcher ,  John A. Howard

发明人： Wesley B. Bruce ,  Elizabeth Hood ,  David J. Peterson ,  James C. Register, III ,  Derrick Witcher ,  John A. Howard

IPC分类号： C12N9/24 ,  C12N15/55 ,  C12N15/82 ,  C12N15/29 ,  A01H4/00 ,  A01H5/00

CPC分类号： C12Y302/01031 ,  C12N15/8242 ,  C12N15/8257 ,  C12N9/2434

摘要： A method for commercial production of GUS 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 GUS-expressing plant. Genetic loci on a plant chromosome are revealed that support high levels of GUS 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GUS的方法需要以天然构象在植物中以表达水平异源表达蛋白质，使得抗生物素蛋白占总提取蛋白质的至少0.1％。 整合基因座的遗传图可以鉴定表达GUS的植物。 显示植物染色体上的遗传基因座，其支持高水平的GUS表达，并且可以用作整合用于其他感兴趣的基因的高水平表达的位点。

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：US06800792B1

公开(公告)日：2004-10-05

申请号：US09786960

申请日：2001-03-12

申请人： John A. Howard ,  Elizabeth Hood ,  Joseph Jilka

发明人： John A. Howard ,  Elizabeth Hood ,  Joseph Jilka

IPC分类号： C12N1509

CPC分类号： C12N9/0061 ,  C12N15/8205 ,  C12N15/8257

摘要： Expression of laccase in plants at commercial levels of production is provided. The laccase gene is preferably operably linked with promoter sequences preferentially directing expression of laccase to the seed of the plant, and may additionally include sequences directing expression to the plant cell wall. Methods of improving the process of introducing DNA into plants via Agrobacterium are also provided.

摘要翻译： 提供漆酶在商业生产水平的植物中的表达。 漆酶基因优选与优先引导漆酶表达的植物的启动子序列可操作地连接，并且还可以包括将表达指导到植物细胞壁的序列。 还提供了通过农杆菌改进将DNA导入植物的过程的方法。

公开(公告)号：US07049484B2

公开(公告)日：2006-05-23

申请号：US10654823

申请日：2003-09-04

申请人： John A. Howard ,  Elizabeth Hood

发明人： John A. Howard ,  Elizabeth Hood

IPC分类号： C12N5/04 ,  C12N15/57 ,  C12N15/82 ,  A01H5/00 ,  A01H5/10

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.

公开(公告)号：US06632930B2

公开(公告)日：2003-10-14

申请号：US09881165

申请日：2001-06-14

申请人： Elizabeth Hood ,  John A. Howard ,  Michele Bailey ,  Franciscus J. C. van Gastel ,  Michael Ward ,  Huaming Wang ,  Susan Woodard

发明人： Elizabeth Hood ,  John A. Howard ,  Michele Bailey ,  Franciscus J. C. van Gastel ,  Michael Ward ,  Huaming Wang ,  Susan Woodard

IPC分类号： A23J100

CPC分类号： C12Y110/03002 ,  C12N9/0061 ,  C12N9/16 ,  C12N9/96 ,  C12N15/8257

摘要： A method of increasing recovery of active enzyme produced in a plant is provided where the enzyme requires a transitional metal cofactor for activation. The metal cofactor is supplied to the enzyme during plant development, during extraction, or after extraction. Recovery of active enzyme is also provided by incubating the extracted enzyme at a non-enzyme degrading temperature. Addition of a negative ion salt further improves active enzyme recovery. Optimum salt concentrations for recovery of laccase from plants using copper solutions is provided.

摘要翻译： 提供了增加植物中产生的活性酶的回收率的方法，其中酶需要用于活化的过渡金属辅因子。 在植物发育，提取期间或提取后，将金属辅因子提供给酶。 通过在非酶降解温度下温育提取的酶也可提供活性酶的回收。 加入负离子盐进一步提高了活性酶的回收率。 提供了使用铜溶液从植物中回收漆酶的最佳盐浓度

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：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。