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1. 发明授权

US09617551B2 Method of increasing plant transformation frequency using modified strains of Agrobacteria 有权
公开(公告)号：US09617551B2
公开(公告)日：2017-04-11
申请号：US13812469
申请日：2011-07-29
申请人： Donald J. Merlo , Sean M. Russell , Diane M. Retallack , Aaron T. Woosley , Thomas Meade , Kenneth Narva
发明人： Donald J. Merlo , Sean M. Russell , Diane M. Retallack , Aaron T. Woosley , Thomas Meade , Kenneth Narva
IPC分类号： C12N15/82 , C12N15/74
CPC分类号： C12N15/8205 , C07K14/195 , C07K14/325 , C12N15/74 , C12N15/743 , C12N15/8274 , C12N15/8286 , Y02A40/162
摘要： Agrobacterium strains that harbor transformation-enhancing genes on a plasmid capable of replication independently of the Agrobacterium chromosome, the Ti plasmid, and plant transformation binary vectors, and uses for these Agrobacterium strains are provided. Additionally, Agrobacterium strains that are deficient in DNA recombination functions that result in instability or rearrangement of plant transformation binary vectors, and that harbor transformation-enhancing genes on a plasmid capable of replication independently of the Agrobacterium chromosome, the Ti plasmid, and plant transformation binary vectors, and uses for these strains, are also provided. Further included are Agrobacterium strains that harbor transformation-enhancing genes integrated into the Agrobacterium chromosome at a locus that does not interfere with or otherwise compromise the normal growth and plant transformation ability of the Agrobacterium cells, and uses for these Agrobacterium strains. Plants made using these Agrobacterium strains are also described.

2. 发明申请

US20150267213A1 STRAINS OF AGROBACTERIUM MODIFIED TO INCREASE PLANT TRANSFORMATION FREQUENCY 有权
标题翻译：修改农杆菌菌株以增加植物转化频率
公开(公告)号：US20150267213A1
公开(公告)日：2015-09-24
申请号：US13812469
申请日：2011-07-29
申请人： Donald J. Merlo , Sean M. Russell , Diane M. Retallack , Aaron T. Woosley , Thomas Meade , Kenneth Narva
发明人： Donald J. Merlo , Sean M. Russell , Diane M. Retallack , Aaron T. Woosley , Thomas Meade , Kenneth Narva
IPC分类号： C12N15/82
CPC分类号： C12N15/8205 , C07K14/195 , C07K14/325 , C12N15/74 , C12N15/743 , C12N15/8274 , C12N15/8286 , Y02A40/162
摘要： Agrobacterium strains that harbor transformation-enhancing genes on a plasmid capable of replication independently of the Agrobacterium chromosome, the Ti plasmid, and plant transformation binary vectors, and uses for these Agrobacterium strains are provided. Additionally, Agrobacterium strains that are deficient in DNA recombination functions that result in instability or rearrangement of plant transformation binary vectors, and that harbor transformation-enhancing genes on a plasmid capable of replication independently of the Agrobacterium chromosome, the Ti plasmid, and plant transformation binary vectors, and uses for these strains, are also provided. Further included are Agrobacterium strains that harbor transformation-enhancing genes integrated into the Agrobacterium chromosome at a locus that does not interfere with or otherwise compromise the normal growth and plant transformation ability of the Agrobacterium cells, and uses for these Agrobacterium strains. Plants made using these Agrobacterium strains are also described.
摘要翻译：提供了在能够独立于农杆菌染色体，Ti质粒和植物转化双向载体复制的质粒上携带转化增强基因的土壤杆菌菌株，以及这些农杆菌菌株的用途。另外，缺乏DNA重组功能的农杆菌菌株导致植物转化双向载体的不稳定或重排，并且能够独立于农杆菌染色体，Ti质粒和植物转化二元体的能够复制的质粒上转录增强基因还提供了这些菌株的载体和用途。还包括农杆菌菌株，其在不干扰或以其他方式损害农杆菌细胞的正常生长和植物转化能力的位点处将整合到农杆菌染色体中的转化增强基因并用于这些农杆菌菌株。还描述了使用这些农杆菌菌株制备的植物。

3. 发明授权

US09394571B2 Method for rapidly screening microbial hosts to identify certain strains with improved yield and/or quality in the expression of heterologous proteins 有权
标题翻译：用于快速筛选微生物宿主以鉴定具有提高异源蛋白质表达的产量和/或质量的某些菌株的方法
公开(公告)号：US09394571B2
公开(公告)日：2016-07-19
申请号：US12109554
申请日：2008-04-25
申请人： Thomas M. Ramseier , Russell J. Coleman , Jane C. Schneider , Charles D. Hershberger , Diane M. Retallack , Charles H. Squires
发明人： Thomas M. Ramseier , Russell J. Coleman , Jane C. Schneider , Charles D. Hershberger , Diane M. Retallack , Charles H. Squires
IPC分类号： G01N33/554 , C12N1/20 , C12N15/00 , C12P21/04 , C12P21/06 , C07H21/04 , C12Q1/68 , C12N15/78
CPC分类号： C12Q1/689 , C12N1/20 , C12N15/78
摘要： The present invention provides an array for rapidly identifying a host cell population capable of producing a heterologous protein with improved yield and/or quality. The array comprises one or more host cell populations that have been genetically modified to increase the expression of one or more target genes involved in protein production, decrease the expression of one or more target genes involved in protein degradation, or both. One or more of the strains in the array may express the heterologous protein of interest in a periplasm compartment or may secrete the heterologous protein extracellularly through an outer cell wall. The strain arrays are useful for screening for improved expression of any protein of interest including therapeutic proteins, hormones, growth factors, extracellular receptors or ligands, proteases, kinases, blood proteins, chemokines, cytokines, antibodies and the like.
摘要翻译：本发明提供了一种用于快速鉴定能够产生具有改善的产量和/或质量的异源蛋白质的宿主细胞群的阵列。该阵列包含已被遗传修饰的一个或多个宿主细胞群，以增加涉及蛋白质产生的一个或多个靶基因的表达，减少参与蛋白质降解的一个或多个靶基因的表达，或两者。阵列中的一个或多个菌株可以在周质隔室中表达感兴趣的异源蛋白质，或者通过外细胞壁在细胞外分泌异源蛋白质。菌株阵列可用于筛选任何目的蛋白质的表达，包括治疗性蛋白质，激素，生长因子，细胞外受体或配体，蛋白酶，激酶，血液蛋白，趋化因子，细胞因子，抗体等。

4. 发明授权

US07794972B2 Benzoate-and anthranilate-inducible promoters 失效
标题翻译：苯甲酸盐和邻氨基苯甲酸盐诱导型启动子
公开(公告)号：US07794972B2
公开(公告)日：2010-09-14
申请号：US11028156
申请日：2005-01-03
申请人： Diane M. Retallack , Venkiteswaran Subramanian
发明人： Diane M. Retallack , Venkiteswaran Subramanian
IPC分类号： C12P21/02 , C12N1/21 , C07H21/04
CPC分类号： C07K14/21 , C12N15/78
摘要： Novel benzoate- or anthranilate-inducible promoters, and novel tandem promoters, and variants and improved mutants thereof, useful for commercial prokaryotic fermentation systems, nucleic acid constructs containing the promoters, expression systems using them, methods for expressing proteins by use thereof, and proteins expressed thereby.
摘要翻译：可用于商业原核发酵系统的新型苯甲酸盐或邻氨基苯甲酸盐诱导型启动子和新型串联启动子及其变体和改良突变体，含有启动子的核酸构建体，使用它们的表达系统，使用它们表达蛋白质的方法和蛋白质由此表达。

5. 发明授权

US09708616B2 Production of recombinant proteins utilizing non-antibiotic selection methods and the incorporation of non-natural amino acids therein 有权
公开(公告)号：US09708616B2
公开(公告)日：2017-07-18
申请号：US14974897
申请日：2015-12-18
申请人： Diane M. Retallack , Lawrence C. Chew , Charles H. Squires
发明人： Diane M. Retallack , Lawrence C. Chew , Charles H. Squires
IPC分类号： C12N15/00 , C12N15/65 , C12P21/02 , C12N15/78 , C12N15/63
CPC分类号： C12N15/65 , C12N15/63 , C12N15/78 , C12P21/02
摘要： Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analog into the growth media for the host cell such that the analog is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analog.

6. 发明授权

US09243253B2 Production of recombinant proteins utilizing non-antibiotic selection methods and the incorporation of non-natural amino acids therein 有权
标题翻译：使用非抗生素选择方法生产重组蛋白，并在其中掺入非天然氨基酸
公开(公告)号：US09243253B2
公开(公告)日：2016-01-26
申请号：US13319844
申请日：2010-05-10
申请人： Diane M. Retallack , Lawrence C. Chew , Charles H. Squires
发明人： Diane M. Retallack , Lawrence C. Chew , Charles H. Squires
IPC分类号： C12N15/00 , C12N15/65 , C12P21/02 , C12N15/78 , C12N15/63
CPC分类号： C12N15/65 , C12N15/63 , C12N15/78 , C12P21/02
摘要： Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analogue into the growth media for the host cell such that the analogue is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analogue.
摘要翻译：本文提供了用于表达包含编码a）重组多肽的核酸的核酸构建体的方法和组合物，以及b）对于至少一种代谢产物是营养缺陷型的宿主细胞中的原始营养恢复酶。在各种实施方案中，宿主细胞对于含氮碱性化合物或氨基酸是营养缺陷型的。本发明涉及将类似物引入宿主细胞的生长培养基中，使得类似物掺入重组多肽或其核酸编码序列中。在各种实施方案中，与抗生素选择系统中的重组蛋白的表达相比，本文公开的组合物和方法导致改善的重组蛋白表达，或与缺乏代谢物类似物的表达系统中重组蛋白的表达进行比较。

7. 发明申请

US20110287443A1 HIGH LEVEL EXPRESSION OF RECOMBINANT TOXIN PROTEINS 有权
标题翻译：重组蛋白的高水平表达
公开(公告)号：US20110287443A1
公开(公告)日：2011-11-24
申请号：US13073955
申请日：2011-03-28
申请人： Diane M. Retallack , Lawrence Chew , Hongfan Jin , Henry W. Talbot
发明人： Diane M. Retallack , Lawrence Chew , Hongfan Jin , Henry W. Talbot
IPC分类号： G01N33/53 , C07K14/195 , C12P21/00
CPC分类号： C12N9/1051 , C07K14/21 , C07K14/235 , C07K14/28 , C07K14/34 , C07K2319/036 , C12N9/1077 , C12N15/78 , C12P21/00 , C12P21/02 , C12Y204/01 , C12Y204/02036 , C12Y304/24068 , G01N33/573
摘要： The present invention relates to the field of recombinant toxin protein production in bacterial hosts. In particular, the present invention relates to production processes for obtaining high levels of a recombinant CRM197, Diphtheria Toxin, Pertussis Toxin, Tetanus Toxoid Fragment C, Cholera Toxin B, Cholera holotoxin, and Pseudomonas Exotoxin A, from a bacterial host.
摘要翻译：本发明涉及细菌宿主中重组毒素蛋白质生产领域。特别地，本发明涉及从细菌宿主获得高水平的重组CRM197，白喉毒素，百日咳毒素，破伤风毒素片段C，霍乱毒素B，霍乱全毒素和假单胞菌外毒素A的生产方法。

8. 发明申请

US20100137162A1 Method for Rapidly Screening Microbial Hosts to Identify Certain Strains with Improved Yield and/or Quality in the Expression of Heterologous Proteins 有权
标题翻译：快速筛选微生物宿主鉴定异源蛋白表达中产量和/或质量提高的某些菌株的方法
公开(公告)号：US20100137162A1
公开(公告)日：2010-06-03
申请号：US12610207
申请日：2009-10-30
申请人： Diane M. Retallack , Charles H. Squires , Thomas M. Ramseier , Russell J. Coleman , Jane C. Schneider , Charies D. Hershberger
发明人： Diane M. Retallack , Charles H. Squires , Thomas M. Ramseier , Russell J. Coleman , Jane C. Schneider , Charies D. Hershberger
IPC分类号： C40B30/10 , C40B50/06 , C40B40/02
CPC分类号： C12N15/1037 , C12N15/78
摘要： The present invention provides an array for rapidly identifying a host cell population capable of producing heterologous protein with improved yield and/or quality. The array comprises one or more host cell populations that have been genetically modified to increase the expression of one or more target genes involved in protein production, decrease the expression of one or more target genes involved in protein degradation, or both. One or more of the strains in the array may express the heterologous protein of interest in a periplasm compartment, or may secrete the heterologous protein extracellularly through an outer cell wall. The strain arrays are useful for screening for improved expression of any protein of interest, including therapeutic proteins, hormones, a growth factors, extracellular receptors or ligands, proteases, kinases, blood proteins, chemokines, cytokines, antibodies and the like.
摘要翻译：本发明提供了一种用于快速鉴定能够产生具有改善的产量和/或质量的异源蛋白质的宿主细胞群的阵列。该阵列包含已被遗传修饰的一个或多个宿主细胞群，以增加涉及蛋白质产生的一个或多个靶基因的表达，减少参与蛋白质降解的一个或多个靶基因的表达，或两者。阵列中的一个或多个菌株可以在周质隔室中表达感兴趣的异源蛋白质，或者通过外细胞壁在细胞外分泌异源蛋白质。菌株阵列可用于筛选任何目的蛋白质的表达，包括治疗性蛋白质，激素，生长因子，细胞外受体或配体，蛋白酶，激酶，血液蛋白，趋化因子，细胞因子，抗体等。

9. 发明授权

US09580719B2 Method for rapidly screening microbial hosts to identify certain strains with improved yield and/or quality in the expression of heterologous proteins 有权
标题翻译：用于快速筛选微生物宿主以鉴定具有提高异源蛋白质表达的产量和/或质量的某些菌株的方法
公开(公告)号：US09580719B2
公开(公告)日：2017-02-28
申请号：US12610207
申请日：2009-10-30
申请人： Diane M. Retallack , Charles H. Squires , Thomas M. Ramseier , Russell J. Coleman , Jane C. Schneider , Charles D. Hershberger
发明人： Diane M. Retallack , Charles H. Squires , Thomas M. Ramseier , Russell J. Coleman , Jane C. Schneider , Charles D. Hershberger
IPC分类号： C12N15/78
CPC分类号： C12N15/1037 , C12N15/78
摘要： The present invention provides an array for rapidly identifying a host cell population capable of producing a heterologous protein with improved yield and/or quality. The array comprises one or more host cell populations that have been genetically modified to increase the expression of one or more target genes involved in protein production, decrease the expression of one or more target genes involved in protein degradation, or both. One or more of the strains in the array may express the heterologous protein of interest in a periplasm compartment or may secrete the heterologous protein extracellularly through an outer cell wall. The strain arrays are useful for screening for improved expression of any protein of interest including therapeutic proteins, hormones, growth factors, extracellular receptors or ligands, proteases, kinases, blood proteins, chemokines, cytokines, antibodies and the like.
摘要翻译：本发明提供了一种用于快速鉴定能够产生具有改善的产量和/或质量的异源蛋白质的宿主细胞群的阵列。该阵列包含已被遗传修饰的一个或多个宿主细胞群，以增加参与蛋白质产生的一个或多个靶基因的表达，减少参与蛋白质降解的一个或多个靶基因的表达，或两者。阵列中的一个或多个菌株可以在周质隔室中表达感兴趣的异源蛋白质，或者通过外细胞壁在细胞外分泌异源蛋白质。菌株阵列可用于筛选任何目的蛋白质的表达，包括治疗性蛋白质，激素，生长因子，细胞外受体或配体，蛋白酶，激酶，血液蛋白，趋化因子，细胞因子，抗体等。

10. 发明申请

US20120115187A1 Production Of Recombinant Proteins Utilizing Non-Antibiotic Selection Methods And The Incorporation Of Non-Natural Amino Acids Therein 有权
标题翻译：生产使用非抗生素选择方法的重组蛋白质和其中非天然氨基酸的掺入
公开(公告)号：US20120115187A1
公开(公告)日：2012-05-10
申请号：US13319844
申请日：2010-05-10
申请人： Diane M. Retallack
发明人： Diane M. Retallack
IPC分类号： C12P21/00
CPC分类号： C12N15/65 , C12N15/63 , C12N15/78 , C12P21/02
摘要： Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analogue into the growth media for the host cell such that the analogue is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analogue.
摘要翻译：本文提供了用于表达包含编码a）重组多肽的核酸的核酸构建体的方法和组合物，以及b）对于至少一种代谢产物是营养缺陷型的宿主细胞中的原始营养恢复酶。在各种实施方案中，宿主细胞对于含氮碱性化合物或氨基酸是营养缺陷型的。本发明涉及将类似物引入宿主细胞的生长培养基中，使得类似物掺入重组多肽或其核酸编码序列中。在各种实施方案中，与抗生素选择系统中的重组蛋白的表达相比，本文公开的组合物和方法导致改善的重组蛋白表达，或与缺乏代谢物类似物的表达系统中重组蛋白的表达进行比较。

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