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公开(公告)号：US6093537A
公开(公告)日：2000-07-25
申请号：US51512
申请日：1993-04-22
申请人： Thomas C. Goodman , Edwin F. Ullman
发明人： Thomas C. Goodman , Edwin F. Ullman
CPC分类号： C12Q1/6804 , C12Q1/6813 , C12Q1/6816 , Y10S435/81 , Y10S530/81
摘要： A method for the detection of a polynucleotide target sequence is described. The method involves the formation of a covalent or non-covalent bonded pair of nucleotide sequences formed in response to a target polynucleotide sequence, adding nucleotide sequence specific binding proteins each capable of binding one member of the pair of nucleotide sequences, and detecting the specific binding proteins complexed to the pair of nucleotide sequences.
摘要翻译： 描述了用于检测多核苷酸靶序列的方法。 所述方法包括形成响应于靶多核苷酸序列形成的共价或非共价键合的核苷酸序列对，加入各自能够结合该对核苷酸序列中的一个成员的核苷酸序列特异性结合蛋白，以及检测特异性结合 与一对核苷酸序列复合的蛋白质。
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公开(公告)号：US5508178A
公开(公告)日：1996-04-16
申请号：US194140
申请日：1994-02-09
CPC分类号： C12Q1/6858 , Y10S435/81
摘要： A method is disclosed for determining the presence of a polynucleotide analyte in a sample suspected of containing the analyte. The method comprises (a) forming as a result of the presence of an analyte a single stranded polynucleotide comprising a target polynucleotide binding sequence flanked by first and second polynucleotide sequences that differ from the sequence of the analyte or a sequence complementary to the analyte sequence, (b) forming multiple copies of the single stranded polynucleotide, and (c) detecting the single stranded polynucleotide. Also disclosed is a method of producing at least one copy of a single stranded polynucleotide. The method comprises (a) forming in the presence of nucleoside triphosphates and template dependent polynucleotide polymerase an extension of a polynucleotide primer at least the 3'-end of which has at least a 10 base sequence hybridizable with a second sequence flanking the 3'-end of the single stranded polynucleotide, the second sequence being partially or fully complementary with at least a 10 base first sequence flanking the 5' end of the single stranded polynucleotide, (b) dissociating the extended polynucleotide primer and the single stranded polynucleotide, (c) repeating step a and (d) dissociating the extended polynucleotide primer and the copy of the single stranded polynucleotide.
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公开(公告)号：US5478729A
公开(公告)日：1995-12-26
申请号：US234456
申请日：1994-04-28
IPC分类号： G01N33/53 , G01N33/543 , G01N33/68 , G01N33/546 , G01N33/577
CPC分类号： G01N33/6815 , G01N33/53 , G01N33/5306 , Y10S435/975 , Y10S436/815 , Y10S436/825
摘要： This invention pertains to methods to detect a compound in the presence of a homolog that is immunologically related to the analyte. The invention is particularly suited for the detection of homocysteine in the presence of cysteine. The methods of this invention involve chemically modifying both the analyte and the homolog to increase their immunogenicity and facilitate antibody recognition. More importantly, this modification is done to make these compounds immunologically distinct. Antibodies to the immunologically distinct compounds are then prepared. An assay protocol comprises chemically modifying the analyte and homolog and then immunochemically detecting the modified analyte by means of the aforementioned antibodies.
摘要翻译： 本发明涉及在与分析物免疫相关的同系物存在下检测化合物的方法。 本发明特别适用于在半胱氨酸存在下检测同型半胱氨酸。 本发明的方法涉及化学修饰分析物和同源物以增加其免疫原性并促进抗体识别。 更重要的是，进行这种修饰以使这些化合物具有免疫学上的不同。 然后制备免疫学上不同化合物的抗体。 测定方案包括化学修饰分析物和同源物，然后通过上述抗体免疫化学检测修饰的分析物。
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4.发明授权公开(公告)号：US06455288B1
公开(公告)日：2002-09-24
申请号：US08044857
申请日：1993-04-08
申请人： Edward Benjamin Jakobovits , Joy L. Silen , Mark J. Levy , Thomas C. Goodman , Martin Becker , Edwin F. Ullman , Robert M. Caldwell , Richard R. Bott , Christopher Charles Barnett
发明人： Edward Benjamin Jakobovits , Joy L. Silen , Mark J. Levy , Thomas C. Goodman , Martin Becker , Edwin F. Ullman , Robert M. Caldwell , Richard R. Bott , Christopher Charles Barnett
IPC分类号： C12N1552
CPC分类号： C12N9/0006 , C12Q1/28 , G01N33/535 , G01N33/56911 , G01N33/581 , G01N33/78 , G01N33/94 , G01N2333/904
摘要： The present invention relates to methods for immunoassay of analytes employing mutant glucose-6-phosphate dehydrogenase (G6PDH) enzymes as labels. In particular, the invention relates-to the use of conjugates of an analyte or analyte analog and a mutant NAD+ dependent G6PDH differing from any precursor G6PDH by the deletion, substitution, or insertion, or any combination thereof of at least one amino acid per subunit. The invention also involves the construction of several mutations in precursor glucose-6-phosphate dehydrogenase (G6PDH) enzymes. Typically, the mutations involve deletion or substitution of one or more lysine residues, or introduction of one or more cysteine residues by insertion of cysteine to precursor G6PDH or substitution of precursor G6PDH amino acids residues with cysteine. The present invention also relates to conjugates of the subject enzymes and specific binding pair members, kits useful in performing the methods of the invention, cell lines producing the subject enzymes, DNA sequences encoding the subject enzymes, and vectors containing DNA encoding the subject enzymes and designed to allow a host cell to produce the subject enzymes.
摘要翻译： 本发明涉及使用突变型葡萄糖-6-磷酸脱氢酶（G6PDH）酶作为标记免疫测定分析物的方法。 特别地，本发明涉及分离物或分析物类似物和与任何前体G6PDH不同的突变体NAD +依赖性G6PDH通过每个亚基的缺失，取代或插入或其任何组合的至少一个氨基酸的缀合物的用途 。 本发明还涉及前体葡萄糖-6-磷酸脱氢酶（G6PDH）酶的几个突变的构建。 通常，突变涉及一个或多个赖氨酸残基的缺失或取代，或通过将半胱氨酸插入前体G6PDH或用半胱氨酸取代前体G6PDH氨基酸残基引入一个或多个半胱氨酸残基。 本发明还涉及本发明的酶和特异性结合对成员的缀合物，可用于实施本发明方法的试剂盒，产生本发明酶的细胞系，编码本发明酶的DNA序列，和含有编码本发明酶的DNA的载体和 旨在使宿主细胞产生目标酶。
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公开(公告)号：US5516641A
公开(公告)日：1996-05-14
申请号：US401660
申请日：1995-03-10
CPC分类号： C07H21/00 , C12Q1/6816 , C12Q1/6834 , C12Q1/6876 , Y10S435/81 , Y10S435/975 , Y10T436/143333
摘要： A kit is disclosed for a method for detecting the presence of a target polynucleotide sequence. The kit comprises a first polynucleotide sequence and a second polynucleotide sequence complementary to non-contiguous portions of a target polynucleotide sequence, which first and second sequences are covalently attached when they are hybridized to the target sequence. The presence of the covalently attached first and second sequences is related to the presence of the target polynucleotide sequence. The invention may be applied to target polynucleotide sequences in DNA or RNA. Specific target polynucleotide sequences of interest will frequently be characteristic of particular microorganisms, viruses, viroids, or genetic characteristics, including genetic abnormalities.
摘要翻译： 公开了用于检测靶多核苷酸序列存在的方法的试剂盒。 该试剂盒包含与靶多核苷酸序列的非连续部分互补的第一多核苷酸序列和第二多核苷酸序列，当与靶序列杂交时，第一和第二序列共价连接。 共价连接的第一和第二序列的存在与靶多核苷酸序列的存在有关。 本发明可以应用于DNA或RNA中的靶多核苷酸序列。 特定目标多核苷酸序列通常是特定微生物，病毒，类病毒或遗传特征的特征，包括遗传异常。
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公开(公告)号：US5185243A
公开(公告)日：1993-02-09
申请号：US236967
申请日：1988-08-25
CPC分类号： C07H21/00 , C12Q1/6816 , C12Q1/6834 , C12Q1/6876 , Y10S435/81 , Y10S435/975 , Y10T436/143333
摘要： A method is disclosed for detecting the presence of a target nucleotide sequence in a polynucleotide. The method comprises hybridizing a first nucleotide sequence and a second nucleotide sequence to non-contiguous portions of a target nucleotide sequence, covalently attaching the first and second sequences when they are hybridized to the target sequence, and determining the presence of covalently attached first and second sequences. The presence of the covalently attached first and second sequences is related to the presence of the target nucleotide sequence. The invention may be applied to target nucleotide sequences in DNA or RNA. Specific target nucleotide sequences of interest will frequently be characteristic of particular microorganisms, viruses, viroids, or genetic characteristics, including genetic abnormalities.
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7.发明授权
公开(公告)号：US6090567A
公开(公告)日：2000-07-18
申请号：US445464
申请日：1995-05-22
申请人： Edward Benjamin Jakobovits , Joy L. Silen , Mark J. Levy , Thomas C. Goodman , Martin Becker , Edwin F. Ullman , Robert M. Caldwell , Richard R. Bott , Christopher Charles Barnett
发明人： Edward Benjamin Jakobovits , Joy L. Silen , Mark J. Levy , Thomas C. Goodman , Martin Becker , Edwin F. Ullman , Robert M. Caldwell , Richard R. Bott , Christopher Charles Barnett
IPC分类号： G01N33/542 , C12N9/04 , C12N15/09 , C12Q1/28 , C12Q1/32 , C12R1/19 , G01N33/535 , G01N33/569 , G01N33/58 , G01N33/78 , G01N33/94 , G01N33/53
CPC分类号： C12N9/0006 , C12Q1/28 , G01N33/535 , G01N33/56911 , G01N33/581 , G01N33/78 , G01N33/94 , G01N2333/904
摘要： Methods for immunoassay of analytes employing mutant glucose-6-phosphate dehydrogenase (G6PDH) enzymes as labels. In particular, the invention relates to the use of conjugates of an analyte or analyte analog and a mutant NAD.sup.+ dependent G6PDH differing from any precursor G6PDH by the deletion, substitution, or insertion, or any combination thereof of at least one amino acid per subunit. The invention also involves the construction of several mutations in precursor glucose-6-phosphate dehydrogenase (G6PDH) enzymes. Typically, the mutations involve deletion or substitution of one or more lysine residues, or introduction of one or more cysteine residues by insertion of cysteine to precursor G6PDH or substitution of precursor G6PDH amino acids residues with cysteine. The present invention also relates to conjugates of the subject enzymes and specific binding pair members, kits useful in performing the methods of the invention, cell lines producing the subject enzymes, DNA sequences encoding the subject enzymes, and vectors containing DNA encoding the subject enzymes and designed to allow a host cell to produce the subject enzymes.
摘要翻译： 使用突变型葡萄糖-6-磷酸脱氢酶（G6PDH）酶作为标记免疫测定分析物的方法。 特别地，本发明涉及通过每个亚基的至少一个氨基酸的缺失，取代或插入或其任何组合，使用不同于任何前体G6PDH的分析物或分析物类似物和突变体NAD +依赖性G6PDH的缀合物的用途。 本发明还涉及前体葡萄糖-6-磷酸脱氢酶（G6PDH）酶的几个突变的构建。 通常，突变涉及一个或多个赖氨酸残基的缺失或取代，或通过将半胱氨酸插入前体G6PDH或用半胱氨酸取代前体G6PDH氨基酸残基引入一个或多个半胱氨酸残基。 本发明还涉及本发明的酶和特异性结合对成员的缀合物，可用于实施本发明方法的试剂盒，产生本发明酶的细胞系，编码本发明酶的DNA序列，和含有编码本发明酶的DNA的载体和 旨在使宿主细胞产生目标酶。
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公开(公告)号：US6063565A
公开(公告)日：2000-05-16
申请号：US451706
申请日：1995-05-26
申请人： Thomas C. Goodman , Edwin F. Ullman
发明人： Thomas C. Goodman , Edwin F. Ullman
CPC分类号： C12Q1/6804 , C12Q1/6813 , C12Q1/6816 , Y10S435/81 , Y10S530/81
摘要： A method for the detection of a polynucleotide target sequence is described. The method involves the formation of a covalent or non-covalent bonded pair of nucleotide sequences formed in response to a target polynucleotide sequence, adding nucleotide sequence specific binding proteins each capable of binding one member of the pair of nucleotide sequences, and detecting the specific binding proteins completed to the pair of nucleotide sequences.
摘要翻译： 描述了用于检测多核苷酸靶序列的方法。 所述方法包括形成响应于靶多核苷酸序列形成的共价或非共价键合的核苷酸序列对，加入各自能够结合该对核苷酸序列中的一个成员的核苷酸序列特异性结合蛋白，以及检测特异性结合 蛋白质完成到一对核苷酸序列。
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公开(公告)号：US5629157A
公开(公告)日：1997-05-13
申请号：US432616
申请日：1995-05-02
申请人： Thomas C. Goodman , Edwin F. Ullman
发明人： Thomas C. Goodman , Edwin F. Ullman
CPC分类号： C12Q1/6804 , C12Q1/6813 , C12Q1/6816 , Y10S435/81 , Y10S530/81
摘要： A method for the detection of a polynucleotide target sequence is described. The method involves the formation of a covalent or non-covalent bonded pair of nucleotide sequences formed in response to a target polynucleotide sequence, adding nucleotide sequence specific binding proteins each capable of binding one member of the pair of nucleotide sequences, and detecting the specific binding proteins complexed to the pair of nucleotide sequences.
摘要翻译： 描述了用于检测多核苷酸靶序列的方法。 该方法涉及形成响应于靶多核苷酸序列而形成的共价或非共价键合对核苷酸序列，加入各自能够结合一对核苷酸序列中的一个成员的核苷酸序列特异性结合蛋白，并检测特异性结合 与一对核苷酸序列复合的蛋白质。
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公开(公告)号：US5273879A
公开(公告)日：1993-12-28
申请号：US614180
申请日：1990-11-13
申请人： Thomas C. Goodman , Martin Becker , Edwin F. Ullman , Samuel Rose
发明人： Thomas C. Goodman , Martin Becker , Edwin F. Ullman , Samuel Rose
CPC分类号： C12Q1/701 , C12Q1/6813 , C12Q1/683 , C12Q1/6844 , C12Q1/6853 , C12Q1/6858
摘要： A Kit is disclosed for a method for producing multiple copies of a primary polynucleotide sequence located at the 3' terminus of a polynucleotide. The method comprises (a) forming in the presence of nucleoside triphosphates and template-dependent polynucleotide polymerase an extension of a primary polynucleotide sequence hybridized with a template sequence of a single stranded pattern polynucleotide comprising two or more template sequences each containing one or more site specific cleavage sequences, (b) cleaving into fragments said extension at cleavable polynucleotide sequences in the presence of means for specifically cleaving said cleavable polynucleotide sequences when said extension is hybridized with said site specific cleavage sequences, (c) dissociating said fragments, (d) hybridizing said fragments with single stranded pattern polynucleotide, and repeating steps (a)-(d). Steps (a)-(d) may be conducted simultaneously or wholly or partially sequentially. The method may be applied in the detection of a polynucleotide analyte in a sample suspected of containing such analyte to facilitate such detection. Also disclosed are compositions for conducting the method of the invention.
摘要翻译： 公开了用于产生位于多核苷酸的3'末端的多核苷酸序列的多个拷贝的方法的试剂盒。 该方法包括（a）在核苷三磷酸和模板依赖性多聚核苷酸聚合酶的存在下形成与单链图案多核苷酸的模板序列杂交的主要多核苷酸序列的延伸，所述单链图案多核苷酸包含两个或更多个模板序列，每个模板序列含有一个或多个位点特异性 裂解序列，（b）当所述延伸与所述位点特异性切割序列杂交时，在存在用于特异性切割所述可切割多核苷酸序列的手段的存在下，在可切割多核苷酸序列处切割成片段，（c）解离所述片段，（d）杂交 所述具有单链图案多核苷酸的片段，以及重复步骤（a） - （d）。 步骤（a） - （d）可以同时或全部或部分顺序进行。 该方法可以应用于怀疑含有此类分析物的样品中的多核苷酸分析物的检测以促进这种检测。 还公开了用于进行本发明方法的组合物。
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