公开(公告)号：US20050181465A1

公开(公告)日：2005-08-18

申请号：US11036761

申请日：2005-01-14

申请人： Mitchell Sanders

发明人： Mitchell Sanders

IPC分类号： C12Q1/04 ,  C12Q1/24 ,  C12Q1/37 ,  G01N33/554 ,  G01N33/569

CPC分类号： C12Q1/04 ,  C12Q1/24 ,  C12Q1/37 ,  G01N2333/96486

摘要： A device and method for detecting the presence or absence of a prokaryotic microorganism are provided, comprising the steps of identifying a protein, such as a microbial-specific protease that characterizes the presence of a specific prokaryotic microbe and thereby provides a marker for that microbe; detecting the protease that is a marker for the presence of a specific prokaryotic microbe by cleaving a substrate when the protease is present; and signaling the presence of that protease when cleavage has occurred. More specifically, the method comprises identifying at least one outer membrane protein or a secreted protein that is unique to a particular microbial pathogen such as for example Listeria monocytogenes and that is substrate specific.

摘要翻译： 提供了用于检测原核生物微生物存在或不存在的装置和方法，包括鉴定表征特定原核生物微生物存在的蛋白质，例如微生物特异性蛋白酶的步骤，从而提供该微生物的标记物; 通过在存在蛋白酶时通过切割底物来检测作为特异性原核生物微生物存在的标记物的蛋白酶; 并且当发生裂解时发信号通知该蛋白酶的存在。 更具体地，该方法包括鉴定至少一种特定微生物病原体（例如单核细胞增生利斯特氏菌）特有的外膜蛋白或分泌蛋白，并且其是底物特异性的。

公开(公告)号：US06861403B2

公开(公告)日：2005-03-01

申请号：US09848780

申请日：2001-05-03

申请人： Mitchell C. Sanders

发明人： Mitchell C. Sanders

IPC分类号： C12N15/09 ,  C07K1/13 ,  C07K1/16 ,  C07K14/435 ,  C07K14/47 ,  C12N15/70 ,  C12P21/02 ,  A61K38/00 ,  C07K1/19

CPC分类号： C07K14/43509 ,  C07K2319/00 ,  C12N15/70

摘要： A method for expressing proteins as a fusion chimera with a domain of p26 or alpha crystallin type proteins to improve the protein stability and solubility when over expressed in bacteria such as E. coli is provided. Genes of interest are cloned into the multiple cloning site of the Vector System just downstream of the p26 or alpha crystallin type protein and a thrombin cleavage site. Protein expression is driven by a strong bacterial promoter (TAC). The expression is induced by the addition of 1 mM IPTG that overcomes the lac repression (lac Iq). The soluble recombinant protein is purified using a fusion tag.

摘要翻译： 提供蛋白质作为具有p26或α晶型蛋白型蛋白质结构域的融合嵌合体的方法，以提高在细菌如大肠杆菌中过表达时的蛋白质稳定性和溶解度。 将感兴趣的基因克隆到p26或α晶状体蛋白型蛋白质下游的凝血酶切割位点的Vector系统的多重克隆位点。 蛋白质表达由强细菌启动子（TAC）驱动。 通过加入克服lac抑制的1mM IPTG诱导表达（lacIq）。 使用融合标签纯化可溶性重组蛋白。

公开(公告)号：US07074757B2

公开(公告)日：2006-07-11

申请号：US11070388

申请日：2005-03-01

申请人： Mitchell C. Sanders

发明人： Mitchell C. Sanders

IPC分类号： C07K1/14 ,  C07K1/10

CPC分类号： C07K14/43509 ,  C07K2319/00 ,  C12N15/70

摘要： A method for expressing proteins as a fusion chimera with a domain of p26 or alpha crystallin type proteins to improve the protein stability and solubility when over expressed in bacteria such as E. coli is provided. Genes of interest are cloned into the multiple cloning site of the Vector System just downstream of the p26 or alpha crystallin type protein and a thrombin cleavage site. Protein expression is driven by a strong bacterial promoter (TAC). The expression is induced by the addition of 1 mM IPTG that overcomes the lac repression (lac Iq). The soluble recombinant protein is purified using a fusion tag.

公开(公告)号：US08039228B2

公开(公告)日：2011-10-18

申请号：US10570075

申请日：2004-09-02

申请人： Gerard J. Colpas ,  Shite Sebastian ,  Mitchell C. Sanders

发明人： Gerard J. Colpas ,  Shite Sebastian ,  Mitchell C. Sanders

IPC分类号： C12Q1/66

CPC分类号： G01N33/54366 ,  C12Q1/04 ,  C12Q1/10 ,  C12Q1/14 ,  C12Q1/34 ,  C12Q1/37 ,  G01N33/542 ,  G01N33/581

摘要： Described herein are zymogens, methods of use for zymogens, and devices that incorporate zymogens. The zymogens include a substrate and an enzyme. The substrate can inhibit the enzyme and is a target for a protein produced by a microorganism. When the substrate is modified by a protein produced by a microorganism, the enzyme is activated. The zymogens can be used to amplify detection assays.

摘要翻译： 本文描述的是酶原，用于酶原的方法和包含酶原的装置。 酶原包括底物和酶。 底物可以抑制酶，并且是由微生物产生的蛋白质的靶标。 当底物被微生物产生的蛋白质修饰时，酶被活化。 酶原可用于扩增检测分析。

公开(公告)号：US20050272864A1

公开(公告)日：2005-12-08

申请号：US11070388

申请日：2005-03-01

申请人： Mitchell Sanders

发明人： Mitchell Sanders

IPC分类号： C12N15/09 ,  C07K1/13 ,  C07K1/16 ,  C07K14/435 ,  C07K14/47 ,  C12N15/70 ,  C12P21/02 ,  C08G63/48 ,  C08G63/91

CPC分类号： C07K14/43509 ,  C07K2319/00 ,  C12N15/70

摘要： A method for expressing proteins as a fusion chimera with a domain of p26 or alpha crystallin type proteins to improve the protein stability and solubility when over expressed in bacteria such as E. coli is provided. Genes of interest are cloned into the multiple cloning site of the Vector System just downstream of the p26 or alpha crystallin type protein and a thrombin cleavage site. Protein expression is driven by a strong bacterial promoter (TAC). The expression is induced by the addition of 1 mM IPTG that overcomes the lac repression (lac Iq). The soluble recombinant protein is purified using a fusion tag.

摘要翻译： 提供蛋白质作为具有p26或α晶型蛋白型蛋白质结构域的融合嵌合体的方法，以提高在细菌如大肠杆菌中过表达时的蛋白质稳定性和溶解度。 将感兴趣的基因克隆到p26或α晶状体蛋白型蛋白质下游的凝血酶切割位点的Vector系统的多重克隆位点。 蛋白质表达由强细菌启动子（TAC）驱动。 通过加入克服lac抑制的1mM IPTG诱导表达（lacI）。 使用融合标签纯化可溶性重组蛋白。