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1.发明授权Protein tag comprising a biotinylation domain and method for increasing solubility and determining folding state 有权包含生物素化结构域的蛋白质标签和增加溶解度并确定折叠状态的方法公开(公告)号：US08999897B2
公开(公告)日：2015-04-07
申请号：US10502581
申请日：2003-01-29
IPC分类号： C40B40/10 , C40B30/10 , C40B40/06 , C40B50/00 , C40B20/04 , C12N9/00 , C12N15/62 , C12N15/67 , C12P21/02
CPC分类号： C12N9/93 , C07K2319/23 , C07K2319/60 , C07K2319/90 , C12N15/62 , C12N15/67 , C12P21/02
摘要： The use of a tag moiety comprising a biotinylation domain, such as biotin carboxyl carrier protein (BCCP), as a protein folding marker and protein solubility enhancer in the orientated surface capture of products of heterologously expressed genes is described. Methods for increasing the solubility of proteins and determining the folded state of a protein are also disclosed. The uses and methods of the invention can be carried out in a multiplexed manner on more than one protein in the formation of libraries. In addition the nucleic acid molecule encoding the biotinylation domain of the tag moiety can be used to increase the proportion of clones in a library that express the protein of interest.
摘要翻译： 描述了在异源表达基因的产物的定向表面捕获中使用包含生物素化结构域的标签部分，例如生物素羧基载体蛋白（BCCP）作为蛋白质折叠标记和蛋白质溶解度增强剂。 还公开了增加蛋白质的溶解度和测定蛋白质的折叠状态的方法。 本发明的用途和方法可以以多种方式在多种形式的文库形成蛋白质中进行。 此外，编码标签部分的生物素化结构域的核酸分子可以用于增加表达所关注蛋白质的文库中克隆的比例。
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公开(公告)号：US20090239761A1
公开(公告)日：2009-09-24
申请号：US12291466
申请日：2008-11-10
申请人： Jonathan Michael Blackburn , Benjamin Leslie James Godber , Darren James Hart , Nicholas A. Bockett , Roland Kozlowski , Michael Dyson
发明人： Jonathan Michael Blackburn , Benjamin Leslie James Godber , Darren James Hart , Nicholas A. Bockett , Roland Kozlowski , Michael Dyson
CPC分类号： C12Q1/00 , B01J2219/00387 , B01J2219/00527 , B01J2219/00605 , B01J2219/00608 , B01J2219/0061 , B01J2219/00612 , B01J2219/00628 , B01J2219/00639 , B01J2219/00691 , B01J2219/00722 , B01J2219/00725 , C12Q1/26 , C12Q1/6837 , C40B30/04 , C40B40/06 , C40B40/10 , C40B60/14 , G01N2333/90209 , G01N2500/00
摘要： The inventors herein describe methods for the production of a functional human, animal, plant or microbe protein arrays and methods to assay for interactions between the proteins on the array with molecules of interest, for example, using such arrays to determine the in vitro metabolite profile of any drug. Such protein arrays can be used, for example, to assay, in a parallel fashion, the protein products of DNA sequences encoding drug metabolizing enzymes (DMES) to obtain a toxicology profile. Also described herein is a novel DME expression and purification strategy using detergents and not requiring an ultra-centrifugation step.
摘要翻译： 本文的发明人描述了用于产生功能性人，动物，植物或微生物蛋白质阵列的方法和用于测定阵列上的蛋白质与感兴趣的分子之间的相互作用的方法，例如使用这样的阵列来确定体外代谢物谱 的任何药物 这样的蛋白质阵列可以用于例如以并行方式测定编码药物代谢酶（DMES）的DNA序列的蛋白质产物以获得毒理学谱。 本文还描述了使用洗涤剂并且不需要超离心步骤的新型DME表达和纯化策略。
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公开(公告)号：US20080132425A1
公开(公告)日：2008-06-05
申请号：US11661762
申请日：2005-09-05
申请人： Darren James Hart
发明人： Darren James Hart
CPC分类号： G01N33/6842 , C12N15/1034 , C12N15/1086 , G01N33/6803 , G01N2500/10
摘要： The present invention relates to methods of screening for expression of a soluble candidate protein within an expression library of candidate proteins. The method involves fusing each candidate protein in the library to a peptide substrate and identifying cells that express soluble candidate protein by detecting enzymatic modification of the peptide substrate.
摘要翻译： 本发明涉及在候选蛋白质的表达文库中筛选可溶性候选蛋白质的表达方法。 该方法包括将文库中的每个候选蛋白融合到肽底物，并通过检测肽底物的酶修饰鉴定表达可溶性候选蛋白的细胞。
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公开(公告)号：US08754012B2
公开(公告)日：2014-06-17
申请号：US11661762
申请日：2005-09-05
申请人： Darren James Hart
发明人： Darren James Hart
CPC分类号： G01N33/6842 , C12N15/1034 , C12N15/1086 , G01N33/6803 , G01N2500/10
摘要： The present invention relates to methods of screening for expression of a soluble candidate protein within an expression library of candidate proteins. The method involves fusing each candidate protein in the library to a peptide substrate and identifying cells that express soluble candidate protein by detecting enzymatic modification of the peptide substrate.
摘要翻译： 本发明涉及在候选蛋白质的表达文库中筛选可溶性候选蛋白质的表达方法。 该方法包括将文库中的每个候选蛋白融合到肽底物，并通过检测肽底物的酶修饰鉴定表达可溶性候选蛋白的细胞。
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