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1.发明申请CARBOXY TERMINAL RESIDUES AS PREDICTORS AND REGULATORS OF PROTEIN STABILITY 有权作为蛋白质稳定性的预测因子和调节因子的羧基末端残基公开(公告)号：US20120107904A1
公开(公告)日：2012-05-03
申请号：US12431315
申请日：2009-04-28
申请人： Brian G. Fox , Yong Chang , Gary A. Wesenberg , Craig A. Bingman
发明人： Brian G. Fox , Yong Chang , Gary A. Wesenberg , Craig A. Bingman
CPC分类号： C12N9/0071 , C07K2319/00 , C07K2319/21 , C07K2319/31 , C07K2319/41 , C07K2319/60 , C12Y114/19001
摘要： Compositions and methods are provided that are useful for predicting and controlling the stability of expressed polypeptides. The compositions and methods may be used to predict and as desired, increase or decrease the stability of proteins recombinantly expressed in mycobacteria, for example DesA3 expressed in Mycobacterium smegmatis. At the C terminus and the penultimate position, substitution to residues with charged side chains, large non-polar side chains, or no side chains can be used to reduce or inhibit the protein degradation. At the antepenultimate position from the C terminus, residues with no side chain or acidic side chains can increase the stability, i.e. reduce or inhibit the protein degradation. The combinational substitution of only the last three residues of polypeptides can make the polypeptides more stable during heterologous expression in mycobacterial hosts.
摘要翻译： 提供了可用于预测和控制表达多肽的稳定性的组合物和方法。 组合物和方法可以用于预测和按需要增加或降低在分枝杆菌中重组表达的蛋白质的稳定性，例如在耻垢分枝杆菌中表达的DesA3。 在C末端和倒数第二的位置，可以使用带有带侧链，大非极性侧链或无侧链的残基取代以降低或抑制蛋白质降解。 在C末端的最初位置，没有侧链或酸性侧链的残基可以增加稳定性，即降低或抑制蛋白质降解。 仅在多肽的最后3个残基的组合取代可以使多肽在分枝杆菌宿主的异源表达期间更稳定。
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公开(公告)号：US06927283B2
公开(公告)日：2005-08-09
申请号：US10001453
申请日：2001-10-22
申请人： Brian G. Fox , Jeffrey A. Haas
发明人： Brian G. Fox , Jeffrey A. Haas
IPC分类号： C07K1/00 , C07K14/00 , C12N1/20 , C12N9/10 , C12N15/00 , C12P21/06 , C12Q1/18 , G01N33/53 , G01N33/533 , G01N33/58 , G01N33/68
CPC分类号： G01N33/6842 , G01N33/533 , G01N33/582 , G01N33/6809
摘要： Disclosed are apo-, holo-, and acylated-acyl carrier proteins modified to have a non-radioactive label covalently bonded to a modified tyrosine residue within the acyl carrier protein. Also disclosed are methods of using the labeled acyl carrier proteins to investigate reaction involving or mediated by acyl carrier proteins. Also disclosed are kits containing the non-radioactively-labeled acyl carrier proteins.
摘要翻译： 公开了修饰为具有与酰基载体蛋白内的修饰的酪氨酸残基共价结合的非放射性标记的apo-，全息和酰化 - 酰基载体蛋白。 还公开了使用标记的酰基载体蛋白来研究涉及或由酰基载体蛋白介导的反应的方法。 还公开了含有非放射性标记的酰基载体蛋白的试剂盒。
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3.发明授权
公开(公告)号：US5190870A
公开(公告)日：1993-03-02
申请号：US600575
申请日：1990-10-08
申请人： John D. Lipscomb , Brian G. Fox
发明人： John D. Lipscomb , Brian G. Fox
CPC分类号： C12N9/0073 , C12P1/00 , C12P17/02 , C12P7/02 , C12Y114/13025
摘要： A purified hydroxylase component of the soluble methane monooxygenase enzyme present in the bacterium Methylosinus trichosporium OB3b is found capable of oxidizing hydrocarbons under aerobic conditions in the presence of suitable reducing agents. The hydroxylase can be reduced by commercial reducing agents, such as sodium dithionite and photo- and electrochemical means when in the presence of electron transport components, such as methyl viologen and proflavin. The hydroxylase component can be obtained with high final specific activity when ferrous iron compounds and cysteine are included in the purification buffers used to extract the hydroxylase from bacterial cells.
摘要翻译： 发现存在于细菌Methylosinus trichosporium OB3b中的可溶性甲烷单加氧酶的纯化羟化酶组分能够在需氧条件下在合适的还原剂存在下氧化烃。 当存在电子转运组分如甲基紫精和黄曲霉素时，羟化酶可通过商业还原剂如连二亚硫酸钠和光电化学手段来还原。 当用于从细菌细胞中提取羟化酶的纯化缓冲液中含有亚铁铁化合物和半胱氨酸时，可以获得高的最终比活性的羟化酶组分。
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公开(公告)号：US4220175A
公开(公告)日：1980-09-02
申请号：US906072
申请日：1978-05-15
CPC分类号： F16K11/165 , Y10T137/7668 , Y10T137/87024 , Y10T137/87692
摘要： An anti-scald apparatus for a tub and shower single control faucet which incorporates a novel cam and cam follower arrangement on the hot and cold water valve means which permits substantially more than 180 degrees of rotation of the control handle. Adjustable temperature stops provide both a comfort stop for normal use and a safety stop which can be reached only by activation of a release button carried by the faucet handle by means of which the comfort stop may be overridden by continuing the rotation of the handle.
摘要翻译： 一种用于浴缸和淋浴单独控制龙头的抗烫伤装置，其在热水和冷水阀装置上结合有新颖的凸轮和凸轮从动装置，其允许控制手柄基本上超过180度的旋转。 可调节的温度停止提供正常使用的舒适停止和仅通过激活由水龙头手柄承载的释放按钮可以达到的安全停止，借助于此可以通过继续旋转手柄来覆盖舒适停止。
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公开(公告)号：US20100304405A1
公开(公告)日：2010-12-02
申请号：US12792156
申请日：2010-06-02
申请人： Brian G. Fox , Nathaniel L. Elsen
发明人： Brian G. Fox , Nathaniel L. Elsen
CPC分类号： C12N15/8261 , C12N15/1079 , Y02A40/146
摘要： Methods for the cell-free identification of polypeptide and polypeptide combinations with utility in biomass transformation, as well as specific novel polypeptides and cell-free systems containing polypeptide combinations discovered by such methods are disclosed.
摘要翻译： 公开了在生物质转化中可用的无细胞鉴定多肽和多肽组合的方法，以及通过这些方法发现的特异性新型多肽和含有多肽组合的无细胞系统。
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公开(公告)号：US07531317B2
公开(公告)日：2009-05-12
申请号：US10997651
申请日：2004-11-24
申请人： Brian G. Fox , Paul G. Blommel
发明人： Brian G. Fox , Paul G. Blommel
CPC分类号： C12Q1/37
摘要： The invention discloses a method of determining protease activity, in real time, using fluorescence polarization technology. In particular, the invention provides vectors and a method for their use, which expresses uncharacterized proteins conjugated to a fluorescence tag, which binds specifically to a fluorescent ligand. Cleavage of the recombinant protein results in a fragment of the expressed peptide and results in a change in fluorescence polarization of the fluorophore. The rate of change in fluorescence polarization can be measured in real time and is equivalent to the rate of protease cleavage.
摘要翻译： 本发明公开了使用荧光偏振技术实时测定蛋白酶活性的方法。 特别地，本发明提供载体及其使用方法，其表达与荧光标记物结合的未特异性蛋白质，其特异性结合荧光配体。 重组蛋白的切割导致表达的肽的片段，并导致荧光团的荧光极化的变化。 荧光偏振的变化率可以实时测量，等同于蛋白酶裂解的速率。
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公开(公告)号：US20080286749A1
公开(公告)日：2008-11-20
申请号：US12100925
申请日：2008-04-10
申请人： Brian G. Fox , Paul G. Blommel
发明人： Brian G. Fox , Paul G. Blommel
摘要： Methods for refining the compositions of bacterial growth media to improve heterologous expression of desired recombinant target genes are provided. Also provided are compositions and culture media obtained using the above methods.
摘要翻译： 提供了细菌生长培养基组合物以改善所需重组靶基因的异源表达的方法。 还提供了使用上述方法获得的组合物和培养基。
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公开(公告)号：US20080199869A1
公开(公告)日：2008-08-21
申请号：US11899080
申请日：2007-09-04
申请人： Brian G. Fox , Yong Chang
发明人： Brian G. Fox , Yong Chang
CPC分类号： C12N9/0083 , C07K2319/21 , C07K2319/50 , C12N9/004
摘要： Mycobacterium tuberculosis proteins and protein compositions that are components of a desaturase complex are provided. The Mycobacterium tuberculosis desaturase complex may include a desaturase and an oxidoreductase. The complex may include the rv3229c and rv3230c gene products of Mycobacterium tuberculosis. Vectors for expressing the desaturase and the oxidoreductase can be packaged together, including a label that indicates their use as a complex for analyzing desaturation of fatty acids. In addition, methods for screening target ligands specific for a desaturase complex are also provided.
摘要翻译： 提供了结核分枝杆菌蛋白质和作为去饱和酶复合物成分的蛋白质组合物。 结核分枝杆菌去饱和酶复合物可包括去饱和酶和氧化还原酶。 复合物可能包括结核分枝杆菌的rv3229c和rv3230c基因产物。 用于表达去饱和酶和氧化还原酶的载体可以包装在一起，包括指示它们用作分析脂肪酸去饱和的复合物的标签。 此外，还提供了用于筛选去饱和酶复合物特异性的靶配体的方法。
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9.发明授权Carboxy terminal residues as predictors and regulators of protein stability 有权羧基末端残基作为蛋白质稳定性的预测因子和调节物公开(公告)号：US08268581B2
公开(公告)日：2012-09-18
申请号：US12431315
申请日：2009-04-28
申请人： Brian G. Fox , Yong Chang , Gary A. Wesenberg , Craig A. Bingman
发明人： Brian G. Fox , Yong Chang , Gary A. Wesenberg , Craig A. Bingman
CPC分类号： C12N9/0071 , C07K2319/00 , C07K2319/21 , C07K2319/31 , C07K2319/41 , C07K2319/60 , C12Y114/19001
摘要： Compositions and methods are provided that are useful for predicting and controlling the stability of expressed polypeptides. The compositions and methods may be used to predict and as desired, increase or decrease the stability of proteins recombinantly expressed in mycobacteria, for example DesA3 expressed in Mycobacterium smegmatis. At the C terminus and the penultimate position, substitution to residues with charged side chains, large non-polar side chains, or no side chains can be used to reduce or inhibit the protein degradation. At the antepenultimate position from the C terminus, residues with no side chain or acidic side chains can increase the stability, i.e. reduce or inhibit the protein degradation. The combinational substitution of only the last three residues of polypeptides can make the polypeptides more stable during heterologous expression in mycobacterial hosts.
摘要翻译： 提供了可用于预测和控制表达多肽的稳定性的组合物和方法。 组合物和方法可以用于预测和按需要增加或降低在分枝杆菌中重组表达的蛋白质的稳定性，例如在耻垢分枝杆菌中表达的DesA3。 在C末端和倒数第二的位置，可以使用带有带侧链，大非极性侧链或无侧链的残基取代以降低或抑制蛋白质降解。 在C末端的最初位置，没有侧链或酸性侧链的残基可以增加稳定性，即降低或抑制蛋白质降解。 仅在多肽的最后3个残基的组合取代可以使多肽在分枝杆菌宿主的异源表达期间更稳定。
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公开(公告)号：US20100330649A1
公开(公告)日：2010-12-30
申请号：US12267825
申请日：2008-11-10
申请人： Brian G. Fox , Pablo Sobrado
发明人： Brian G. Fox , Pablo Sobrado
CPC分类号： C07K14/80 , C07K2319/03 , C07K2319/04
摘要： Expression systems and methods for the expression of functional membrane polypeptides such as human cytochrome b5 are provided. The systems include recombinant expression vectors capable of expressing soluble fusion proteins that include a solubilizing agent, a linker, and a membrane polypeptide, as well as one or more cleavers, e.g. proteases, capable of cleaving the linker to release the membrane polypeptide. When the fusion protein is expressed, the linker is cleaved by the cleaver to allow association of the membrane polypeptide with a membrane.
摘要翻译： 提供了用于表达功能性膜多肽如人细胞色素b5的表达系统和方法。 该系统包括能够表达可溶性融合蛋白的重组表达载体，其包括增溶剂，接头和膜多肽，以及一个或多个切割剂，例如， 蛋白酶，能够切割接头以释放膜多肽。 当融合蛋白被表达时，接头被切割器切割以允许膜多肽与膜结合。
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