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公开(公告)号:US06867014B2
公开(公告)日:2005-03-15
申请号:US10651183
申请日:2003-08-28
申请人: Glenn Kawasaki , Heather Kay Webb , Jeffrey Owens , Raymond Liedtke , Doreen Forest , Mark Legaz , Sobomabo Lawson
发明人: Glenn Kawasaki , Heather Kay Webb , Jeffrey Owens , Raymond Liedtke , Doreen Forest , Mark Legaz , Sobomabo Lawson
IPC分类号: C12Q1/00 , C12N9/00 , C12N15/52 , C12Q1/26 , C12Q1/32 , C12Q1/527 , G01N33/53 , G01N33/68 , C12Q1/34 , C12Q1/48
CPC分类号: C12Q1/26 , C12Q1/32 , C12Q1/527 , G01N33/6815 , Y10S435/968 , Y10S435/975
摘要: The present invention provides an enzymatic cycling assay for assessing the amount of homocysteine and/or cystathionine in a solution such as blood, blood derivatives, or urine. The assay comprises the steps of contacting the solution containing homocysteine and/or cystathionine to form a reaction mixture, with CBS, or a derivative thereof, L-serine, and CBL, or a derivative thereof, for a time period sufficient to catalyze the cyclical conversion of homocysteine form to cystathionine and the reconversion of cystathionine to homocysteine with the production of pyruvate and ammonia; determining the amount of homocysteine and/or ammonia present in the reaction mixture; and determining the amount of homocysteine and/or cystathionine present in the solution based on the amount of pyruvate and/or ammonia formed. Expression vectors and isolation procedures for CBS, or derivatives thereof, and CBL, or derivatives thereof, are also provided as well as test kits for carrying out the assay. In preferred embodiments, the assays can be conducted in 15 minutes or less, with a minimum of enzyme usage.
摘要翻译: 本发明提供了用于评估溶液如血液,血液衍生物或尿液中高半胱氨酸和/或胱硫醚的量的酶循环测定。 该测定包括以下步骤:使含有同型半胱氨酸和/或胱硫醚的溶液与CBS或其衍生物,L-丝氨酸和CBL或其衍生物反应一段足以催化周期性 同型半胱氨酸形式转化为胱硫醚,并产生丙酮酸和氨,将胱硫醚转化为同型半胱氨酸; 测定反应混合物中存在的同型半胱氨酸和/或氨的量; 并且基于形成的丙酮酸盐和/或氨的量确定溶液中存在的同型半胱氨酸和/或胱硫醚的量。 还提供用于CBS或其衍生物,CBL或其衍生物的表达载体和分离程序以及用于进行测定的测试试剂盒。 在优选的实施方案中,测定可以在15分钟或更短时间内进行,酶用量最小。
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公开(公告)号:US06664073B1
公开(公告)日:2003-12-16
申请号:US09704036
申请日:2000-11-01
申请人: Glenn Kawasaki , Heather Kay Webb , Jeffrey Owens , Raymond Liedtke , Doreen Forest , Mark Legaz , Sobomabo Lawson
发明人: Glenn Kawasaki , Heather Kay Webb , Jeffrey Owens , Raymond Liedtke , Doreen Forest , Mark Legaz , Sobomabo Lawson
IPC分类号: C12Q126
CPC分类号: C12Q1/26 , C12Q1/32 , C12Q1/527 , Y10S435/968 , Y10S435/975
摘要: The present invention provides an enzymatic cycling assay for assessing the amount of homocysteine and/or cystathionine in a solution such as blood, blood, derivatives, or urine. The assay comprises the steps of contacting the solution containing homocysteine and/or cystathionine to form a reaction mixture, with CBS, or a derivative thereof, L-serine, and CBL, or a derivative thereof, for a time period sufficient to catalyze the cyclical conversion of homocysteine form to cystathionine and the reconversion of cystathionine to homocysteine with the production of pyruvate and ammonia; determining the amount of homocysteine and/or ammonia present in the reaction mixture; and determining the amount of homocysteine and/or cystathionine present in the solution based on the amount of pyruvate and/or ammonia formed. Expression vectors and isolation procedures for CBS, or derivatives thereof, and CBL, or derivatives thereof, are also provided as well as test kits for carrying out the assay. In preferred embodiments, the assays can be conducted in 15 minutes or less, with a minimum of enzyme usage.
摘要翻译: 本发明提供了用于评估溶液如血液,血液,衍生物或尿液中的同型半胱氨酸和/或胱硫醚的量的酶循环试验。 该测定包括以下步骤:使含有同型半胱氨酸和/或胱硫醚的溶液与CBS或其衍生物,L-丝氨酸和CBL或其衍生物反应一段足以催化周期性 同型半胱氨酸形式转化为胱硫醚,并产生丙酮酸和氨,将胱硫醚转化为同型半胱氨酸; 确定反应混合物中存在的同型半胱氨酸和/或氨的量; 并且基于形成的丙酮酸盐和/或氨的量确定溶液中存在的同型半胱氨酸和/或胱硫醚的量。 还提供用于CBS或其衍生物,CBL或其衍生物的表达载体和分离程序以及用于进行测定的测试试剂盒。 在优选的实施方案中,测定可以在15分钟或更短时间内进行,酶用量最小。
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公开(公告)号:US06635438B2
公开(公告)日:2003-10-21
申请号:US10012762
申请日:2001-11-06
申请人: Glenn Kawasaki , Heather Kay Webb , Jeffrey Owens , Doreen Forest , Raymond Liedtke , Sobomabo Lawson , Mark Legaz
发明人: Glenn Kawasaki , Heather Kay Webb , Jeffrey Owens , Doreen Forest , Raymond Liedtke , Sobomabo Lawson , Mark Legaz
IPC分类号: C12Q126
CPC分类号: C12Q1/26 , C12Q1/32 , C12Q1/527 , G01N33/6815 , Y10S435/968 , Y10S435/975
摘要: The present invention provides an enzymatic cycling assay for assessing the amount of homocysteine and/or cystathionine in a solution such as blood, blood derivatives, or urine. The assay comprises the steps of contacting the solution containing homocysteine and/or cystathionine to form a reaction mixture, with CBS, or a derivative thereof, L-serine, and CBL, or a derivative thereof, for a time period sufficient to catalyze the cyclical conversion of homocysteine form to cystathionine and the reconversion of cystathionine to homocysteine with the production of pyruvate and ammonia; determining the amount of homocysteine and/or ammonia present in the reaction mixture; and determining the amount of homocysteine and/or cystathionine present in the solution based on the amount of pyruvate and/or ammonia formed. Expression vectors and isolation procedures for CBS, or derivatives thereof, and CBL, or derivatives thereof, are also provided as well as test kits for carrying out the assay. In preferred embodiments, the assays can be conducted in 15 minutes or less, with a minimum of enzyme usage.
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公开(公告)号:US5700643A
公开(公告)日:1997-12-23
申请号:US473012
申请日:1995-06-06
申请人: Glenn Kawasaki
发明人: Glenn Kawasaki
IPC分类号: C12N15/09 , C07H21/04 , C07K14/15 , C07K14/395 , C07K14/49 , C07K14/81 , C12N1/16 , C12N1/19 , C12N5/00 , C12N9/90 , C12N15/00 , C12N15/64 , C12N15/67 , C12N15/81 , C12P19/34 , C12P21/00 , C12P21/02 , C12R1/645 , C12R1/865
CPC分类号: C07K14/005 , C07K14/395 , C07K14/49 , C07K14/8125 , C12N15/64 , C12N15/67 , C12N15/81 , C12N9/90 , C12N2740/10022 , C12N2740/11022
摘要: Methods are provided for producing protein products in host cells and for selecting transformed cells comprising the step of transforming the host cell with a DNA molecule comprising a gene which complements a deficiency in the host cell. The host cell is a strain having a deficiency in a function necessary for normal cell growth. The gene in the DNA molecule, such as a plasmid, which complements the deficiency serves as a selection marker whereby the growth conditions for selection may comprise a conventional complex medium.
摘要翻译: 提供了用于在宿主细胞中产生蛋白质产物和选择转化细胞的方法,所述细胞包括用包含补体宿主细胞缺陷的基因的DNA分子转化宿主细胞的步骤。 宿主细胞是具有正常细胞生长所必需的功能缺陷的菌株。 补充缺陷的DNA分子中的诸如质粒的基因用作选择标记,由此选择的生长条件可以包含常规的复合培养基。
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公开(公告)号:US4711848A
公开(公告)日:1987-12-08
申请号:US709382
申请日:1985-03-07
申请人: Margaret Y. Insley , Glenn Kawasaki
发明人: Margaret Y. Insley , Glenn Kawasaki
IPC分类号: A61K35/74 , A61K38/00 , A61K38/16 , A61K38/55 , A61P7/02 , A61P43/00 , C07H21/04 , C07K14/005 , C07K14/195 , C07K14/81 , C12N9/99 , C12N15/00 , C12N15/09 , C12N15/10 , C12N15/15 , C12N15/81 , C12P21/02 , C12R1/865 , C12P19/34 , C12N1/00
CPC分类号: C12N15/00 , C07K14/8125 , C12N15/102 , C12N15/81 , A61K38/00
摘要: Methods are provided for producing sites specific mutagenized alpha-1-antitrypsin. Particular mutants are formed having a mutation in the active site of alpha-1-antitrypsin at amino acid position 358 and at amino acid position 342.
摘要翻译: 提供了用于产生位点特异性诱变的α-1-抗胰蛋白酶的方法。 在氨基酸位置358和氨基酸位置342处形成在α-1-抗胰蛋白酶的活性位点具有突变的特异性突变体。
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公开(公告)号:US5527668A
公开(公告)日:1996-06-18
申请号:US134480
申请日:1993-10-12
申请人: Glenn Kawasaki , Leslie Bell
发明人: Glenn Kawasaki , Leslie Bell
IPC分类号: C12N15/09 , C07H21/04 , C07K14/15 , C07K14/395 , C07K14/49 , C07K14/81 , C12N1/16 , C12N1/19 , C12N5/00 , C12N9/90 , C12N15/00 , C12N15/64 , C12N15/67 , C12N15/81 , C12P19/34 , C12P21/00 , C12P21/02 , C12R1/645 , C12R1/865 , C12Q1/68
CPC分类号: C07K14/005 , C07K14/395 , C07K14/49 , C07K14/8125 , C12N15/64 , C12N15/67 , C12N15/81 , C12N9/90 , C12N2740/10022 , C12N2740/11022
摘要: Methods are provided for producing protein products in host cells and for selecting transformed cells comprising the step of transforming the host cell with a DNA molecule comprising a gene which complements a deficiency in the host cell. The host cell is a strain having a deficiency in a function necessary for normal cell growth. The gene in the DNA molecule, such as a plasmid, which complements the deficiency serves as a selection marker whereby the growth conditions for selection may comprise a conventional complex medium.
摘要翻译: 提供了用于在宿主细胞中产生蛋白质产物和选择转化细胞的方法,所述细胞包括用包含补体宿主细胞缺陷的基因的DNA分子转化宿主细胞的步骤。 宿主细胞是具有正常细胞生长所必需的功能缺陷的菌株。 补充缺陷的DNA分子中的诸如质粒的基因用作选择标记,由此选择的生长条件可以包含常规的复合培养基。
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7.
公开(公告)号:US4931373A
公开(公告)日:1990-06-05
申请号:US663315
申请日:1984-10-22
申请人: Glenn Kawasaki , Leslie Bell
发明人: Glenn Kawasaki , Leslie Bell
IPC分类号: C07K14/15 , C07K14/49 , C07K14/81 , C12N9/90 , C12N15/15 , C12N15/64 , C12N15/67 , C12N15/81
CPC分类号: C07K14/005 , C07K14/49 , C07K14/8125 , C12N15/64 , C12N15/67 , C12N15/81 , C12N9/90 , C12N2740/10022
摘要: Methods are provided for producing .alpha.-1-antitrypsin in host cells and for selecting transformed cells comprising the step of transforming the host cell with a DNA molecule comprising a gene which complements a deficiency in the host cell. The host cell is a strain having a deficiency in a function necessary for normal cell growth. The gene in the DNA molecule, such as a plasmid, which complements the deficiency serves as a selectable marker whereby the growth conditions for selection may comprise a conventional complex medium.
摘要翻译: 提供了用于在宿主细胞中产生α-1-抗胰蛋白酶的方法,并用于选择转化细胞,所述转化细胞包括用包含补体宿主细胞缺陷的基因的DNA分子转化宿主细胞的步骤。 宿主细胞是具有正常细胞生长所必需的功能缺陷的菌株。 补充缺陷的DNA分子中的基因(例如质粒)用作可选择标记,由此选择的生长条件可以包含常规的复合培养基。
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