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1.发明申请公开(公告)号：US20100136621A1
公开(公告)日：2010-06-03
申请号：US12692665
申请日：2010-01-25
申请人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
发明人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/50 , C12N9/90
摘要： Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.
摘要翻译： 公开了用于产生不溶性融合肽的肽标签，这里称为包涵体标签。 融合肽包含可操作地连接到目的肽的至少一种包涵体标签。 融合肽在宿主细胞中的表达产生不溶性并且包含在细胞和/或细胞裂解物中的包涵体内的产物。 然后可以纯化包涵体，并且在从包涵体标签中切割后可以分离感兴趣的蛋白质。
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2.发明申请公开(公告)号：US20090029412A1
公开(公告)日：2009-01-29
申请号：US11782836
申请日：2007-07-25
申请人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss
发明人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss
CPC分类号： C12P21/02 , C07K2319/50 , C12N9/90
摘要： Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.
摘要翻译： 公开了用于产生不溶性融合肽的肽标签，这里称为包涵体标签。 融合肽包含可操作地连接到目的肽的至少一种包涵体标签。 融合肽在宿主细胞中的表达产生不溶性并且包含在细胞和/或细胞裂解物中的包涵体内的产物。 然后可以纯化包涵体，并且在从包涵体标签切割后可以分离感兴趣的蛋白质。
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公开(公告)号：US07794979B2
公开(公告)日：2010-09-14
申请号：US12692665
申请日：2010-01-25
申请人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
发明人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/50 , C12N9/90
摘要： Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.
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4.发明授权公开(公告)号：US07678883B2
公开(公告)日：2010-03-16
申请号：US11782836
申请日：2007-07-25
申请人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
发明人： Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Lisa Diane Reiss , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/50 , C12N9/90
摘要： Peptide tags, referred to here as inclusion body tags, are disclosed useful for the generation of insoluble fusion peptides. The fusion peptides comprise at least one inclusion body tag operably linked to a peptide of interest. Expression of the fusion peptide in a host cell results in a product that is insoluble and contained within inclusion bodies in the cell and/or cell lysate. The inclusion bodies may then be purified and the protein of interest may be isolated after cleavage from the inclusion body tag.
摘要翻译： 公开了用于产生不溶性融合肽的肽标签，这里称为包涵体标签。 融合肽包含可操作地连接到目的肽的至少一种包涵体标签。 融合肽在宿主细胞中的表达产生不溶性并且包含在细胞和/或细胞裂解物中的包涵体内的产物。 然后可以纯化包涵体，并且在从包涵体标签中切割后可以分离感兴趣的蛋白质。
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公开(公告)号：US20090043075A1
公开(公告)日：2009-02-12
申请号：US12172395
申请日：2008-07-14
申请人： Albert W. Alsop , Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
发明人： Albert W. Alsop , Qiong Cheng , Linda Jane Decarolis , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/00 , C12P21/06
摘要： The invention relates to the recombinant expression of a peptide of interest in the form of a fusion protein comprising a solubility tag. The fusion protein comprises at least two portions separated by a cleavable peptide sequence wherein one portion is devoid of cysteine residues and the second portion comprises an effective number of cross-linkable cysteine residues. After cell lysis and isolation of the fusion protein, the fusion protein is subsequently cleaved into a mixture of first and second portions. Oxidative cross-linking is used to selectively precipitate one of the two portions to facilitate simple and effective separation of the peptide of interest.
摘要翻译： 本发明涉及以包含溶解性标签的融合蛋白形式的目标肽的重组表达。 融合蛋白包含由可切割肽序列分开的至少两个部分，其中一部分缺少半胱氨酸残基，第二部分包含有效数目的可交联半胱氨酸残基。 细胞裂解和分离融合蛋白后，融合蛋白随后被切割成第一和第二部分的混合物。 氧化交联用于选择性沉淀两部分之一，以促进目的肽的简单和有效分离。
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公开(公告)号：US07951559B2
公开(公告)日：2011-05-31
申请号：US12172395
申请日：2008-07-14
申请人： Albert W. Alsop , Qiong Cheng , Linda Jane Solomon , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
发明人： Albert W. Alsop , Qiong Cheng , Linda Jane Solomon , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/00 , C12P21/06
摘要： The invention relates to the recombinant expression of a peptide of interest in the form of a fusion protein comprising a solubility tag. The fusion protein comprises at least two portions separated by a cleavable peptide sequence wherein one portion is devoid of cysteine residues and the second portion comprises an effective number of cross-linkable cysteine residues. After cell lysis and isolation of the fusion protein, the fusion protein is subsequently cleaved into a mixture of first and second portions. Oxidative cross-linking is used to selectively precipitate one of the two portions to facilitate simple and effective separation of the peptide of interest.
摘要翻译： 本发明涉及以包含溶解性标签的融合蛋白形式的目标肽的重组表达。 融合蛋白包含由可切割肽序列分开的至少两个部分，其中一部分缺少半胱氨酸残基，第二部分包含有效数目的可交联半胱氨酸残基。 在融合蛋白的细胞溶解和分离后，随后将融合蛋白切割成第一和第二部分的混合物。 氧化交联用于选择性沉淀两部分之一，以促进目的肽的简单和有效分离。
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7.发明申请USE OF TETRACYSTEINE TAGS IN FLUORESCENCE-ACTIVATED CELL SORTING ANALYSIS OF PROKARYOTIC CELLS PRODUCING PEPTIDES OR PROTEINS 失效在生产肽或蛋白质的原核细胞的荧光激活细胞分选中使用四肽蛋白标签公开(公告)号：US20090117609A1
公开(公告)日：2009-05-07
申请号：US12263608
申请日：2008-11-03
申请人： Qiong Cheng , Kevin Michael Croker , Stephen R. Fahnestock , Tanja Maria Gruber , Kristin Ruebling-Jass , Jianzhong Zhang
发明人： Qiong Cheng , Kevin Michael Croker , Stephen R. Fahnestock , Tanja Maria Gruber , Kristin Ruebling-Jass , Jianzhong Zhang
CPC分类号： C12N15/62 , C07K2319/20 , C07K2319/60
摘要： A process of in vivo labeling and identifying recombinantly produced peptides or proteins within an unpermeabilized prokaryotic host cell. Recombinant prokaryotic cells expressing a fusion peptide comprising at least one tetracysteine tag were labeled in vivo using a biarsenical labeling reagent. A fluorescent activated cell sorter was used to identify and select subpopulations of fluorescent cells wherein the amount of fusion peptide in the cell was proportional to the amount of fluorescence detected.
摘要翻译： 体内标记和鉴定重组产生的肽或蛋白质在未经过预处理的原核宿主细胞内的过程。 表达含有至少一个四羧酸标签的融合肽的重组原核细胞在体内用双标记标记试剂进行标记。 使用荧光活化细胞分选器来鉴定和选择荧光细胞亚群，其中细胞中融合肽的量与所检测的荧光量成比例。
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公开(公告)号：US08617843B2
公开(公告)日：2013-12-31
申请号：US13080917
申请日：2011-04-06
申请人： Albert W. Alsop , Qiong Cheng , Linda Jane Solomon , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
发明人： Albert W. Alsop , Qiong Cheng , Linda Jane Solomon , Stephen R. Fahnestock , Tanja Maria Gruber , Pierre E. Rouviere
CPC分类号： C12P21/02 , C07K2319/00 , C12P21/06
摘要： The invention relates to the recombinant expression of a peptide of interest in the form of a fusion protein comprising a solubility tag. The fusion protein comprises at least two portions separated by a cleavable peptide sequence wherein one portion is devoid of cysteine residues and the second portion comprises an effective number of cross-linkable cysteine residues. After cell lysis and isolation of the fusion protein, the fusion protein is subsequently cleaved into a mixture of first and second portions. Oxidative cross-linking is used to selectively precipitate one of the two portions to facilitate simple and effective separation of the peptide of interest.
摘要翻译： 本发明涉及以包含溶解性标签的融合蛋白形式的目标肽的重组表达。 融合蛋白包含由可切割肽序列分开的至少两个部分，其中一部分缺少半胱氨酸残基，第二部分包含有效数目的可交联半胱氨酸残基。 细胞裂解和分离融合蛋白后，融合蛋白随后被切割成第一和第二部分的混合物。 氧化交联用于选择性沉淀两部分之一，以促进目的肽的简单和有效分离。
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9.发明授权Use of tetracysteine tags in fluorescence-activated cell sorting analysis of prokaryotic cells producing peptides or proteins 失效在用于产生肽或蛋白质的原核细胞的荧光激活细胞分选分析中使用四羧酸标签公开(公告)号：US07794963B2
公开(公告)日：2010-09-14
申请号：US12263608
申请日：2008-11-03
申请人： Qiong Cheng , Kevin Michael Croker , Stephen R. Fahnestock , Tanja Maria Gruber , Kristin Ruebling-Jass , Jianzhong Zhang
发明人： Qiong Cheng , Kevin Michael Croker , Stephen R. Fahnestock , Tanja Maria Gruber , Kristin Ruebling-Jass , Jianzhong Zhang
IPC分类号： G01N33/569 , C07K16/00
CPC分类号： C12N15/62 , C07K2319/20 , C07K2319/60
摘要： A process of in vivo labeling and identifying recombinantly produced peptides or proteins within an unpermeabilized prokaryotic host cell. Recombinant prokaryotic cells expressing a fusion peptide comprising at least one tetracysteine tag were labeled in vivo using a biarsenical labeling reagent. A fluorescent activated cell sorter was used to identify and select subpopulations of fluorescent cells wherein the amount of fusion peptide in the cell was proportional to the amount of fluorescence detected.
摘要翻译： 体内标记和鉴定重组产生的肽或蛋白质在未经过预处理的原核宿主细胞内的过程。 表达含有至少一个四羧酸标签的融合肽的重组原核细胞在体内用双标记标记试剂进行标记。 使用荧光活化细胞分选器来鉴定和选择荧光细胞亚群，其中细胞中融合肽的量与所检测的荧光量成比例。
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10.发明申请Identification of peptide tags for the production of insoluble peptides by sequence scanning 审中-公开通过序列扫描鉴定用于产生不溶性肽的肽标签公开(公告)号：US20100234568A1
公开(公告)日：2010-09-16
申请号：US11641981
申请日：2006-12-19
CPC分类号： C12N15/1093 , C12N15/1065 , C40B40/08 , C40B50/06
摘要： A method is provided to identify short peptide tags, referred to here as inclusion body tags (IBTs), useful for the generation of insoluble fusion peptides. A library of genetic constructs were prepared encoding fusion peptides comprising an inclusion body tag of 10-50 contiguous amino acids from a full-length insoluble protein operably linked to a peptide of interest. The library was designed to include a sufficient number of overlapping inclusion body tags to ensure that the entire length of the full-length insoluble protein was represented. Host cells transformed and expressing the genetic constructs were evaluated for inclusion body formation.
摘要翻译： 提供了一种用于鉴定可用于产生不溶性融合肽的短肽标签（本文称为包含体标签（IBT））的方法。 制备遗传构建体文库，编码融合肽，其包含可操作地连接到感兴趣肽的全长不溶性蛋白质的10-50个连续氨基酸的包涵体标签。 该文库旨在包括足够数量的重叠包含体标签，以确保全长不溶性蛋白质的全长被表示。 评估转化并表达遗传构建体的宿主细胞的包涵体形成。
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